Pyrrolopyrimidine compounds and their uses

ABSTRACT

The disclosed compounds relate to treatments and therapies for protein kinase-associated disorders. There is also a need for compounds useful in the treatment or prevention or amelioration of one or more symptoms of cancer, transplant rejections, and autoimmune diseases. Furthermore, there is a need for methods for modulating the activity of protein kinases, such as CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9, using the compounds provided herein.

BACKGROUND

The search for new therapeutic agents has been greatly aided in recentyears by a better understanding of the structure of enzymes and otherbiomolecules associated with diseases. One important class of enzymesthat has been the subject of extensive study is protein kinases.

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a variety of signaltransduction processes within the cell. (Hardie, G. and Hanks, S. TheProtein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.:1995). Protein kinases are thought to have evolved from a commonancestral gene due to the conservation of their structure and catalyticfunction. Almost all kinases contain a similar 250-300 amino acidcatalytic domain. The kinases may be categorized into families by thesubstrates they phosphorylate (e.g., protein-tyrosine,protein-serine/threonine, lipids, etc.). Sequence motifs have beenidentified that generally correspond to each of these kinase families(See, for example, Hanks, S. K., Hunter, T., FASEB J. 1995, 9, 576-596;Knighton et al., Science 1991, 253, 407-414; Hiles et al., Cell 1992,70, 419-429; Kunz et al., Cell 1993, 73, 585-596; Garcia-Bustos et al.,EMBO J. 1994, 13, 2352-2361).

In general, protein kinases mediate intracellular signaling by affectinga phosphoryl transfer from a nucleoside triphosphate to a proteinacceptor that is involved in a signaling pathway. These phosphorylationevents act as molecular on/off switches that can modulate or regulatethe target protein biological function. These phosphorylation events areultimately triggered in response to a variety of extracellular and otherstimuli. Examples of such stimuli include environmental and chemicalstress signals (e.g., osmotic shock, heat shock, ultraviolet radiation,bacterial endotoxin, and H₂O₂), cytokines (e.g., interleukin-1 (IL-1)and tumor necrosis factor-α (TNF-α)), and growth factors (e.g.,granulocyte macrophage-colony-stimulating factor (GM-CSF), andfibroblast growth factor (FGF)). An extracellular stimulus may affectone or more cellular responses related to cell growth, migration,differentiation, secretion of hormones, activation of transcriptionfactors, muscle contraction, glucose metabolism, control of proteinsynthesis, and regulation of the cell cycle. Many diseases areassociated with abnormal cellular responses triggered by proteinkinase-mediated events as described above. These diseases include, butare not limited to, autoimmune diseases, inflammatory diseases, bonediseases, metabolic diseases, neurological and neurodegenerativediseases, cancer, cardiovascular diseases, allergies and asthma,Alzheimer's disease, and hormone-related diseases. Accordingly, therehas been a substantial effort in medicinal chemistry to find proteinkinase inhibitors that are effective as therapeutic agents.

Initiation, progression, and completion of the mammalian cell cycle areregulated by various cyclin-dependent kinase (CDK) complexes, which arecritical for cell growth. These complexes comprise at least a catalytic(the CDK itself) and a regulatory (cyclin) subunit. Some of the moreimportant complexes for cell cycle regulation include cyclin A(CDK1—also known as cdc2, and CDK2), cyclin B1-B3 (CDK1) and cyclinD1-D3 (CDK2, CDK4, CDK5, CDK6), cyclin E (CDK2). Each of these complexesis involved in a particular phase of the cell cycle. Not all members ofthe CDK family are involved exclusively in cell cycle control, however.Thus CDKs 7, 8, and 9 are implicated in the regulation of transcription,and CDK5 plays a role in neuronal and secretory cell function.

The activity of CDKs is regulated post-translationally, by transitoryassociations with other proteins, and by alterations of theirintracellular localization. Tumor development is closely associated withgenetic alteration and deregulation of CDKs and their regulators,suggesting that inhibitors of CDKs may be useful anti-cancertherapeutics. Indeed, early results suggest that transformed and normalcells differ in their requirement for, e.g., cyclin A/CDK2 and that itmay be possible to develop novel antineoplastic agents devoid of thegeneral host toxicity observed with conventional cytotoxic andcytostatic drugs. While inhibition of cell cycle-related CDKs is clearlyrelevant in, e.g., oncology applications, this may not be the case forthe inhibition of RNA polymerase-regulating CDKs. On the other hand,inhibition of CDK9/cyclin T function was recently linked to preventionof HIV replication and the discovery of new CDK biology thus continuesto open up new therapeutic indications for CDK inhibitors (Sausville, E.A. Trends Molec. Med. 2002, 8, S32-S37).

The function of CDKs is to phosphorylate and thus activate or deactivatecertain proteins, including e.g. retinoblastoma proteins, lamins,histone H1, and components of the mitotic spindle. The catalytic stepmediated by CDKs involves a phospho-transfer reaction from ATP to themacromolecular enzyme substrate. Several groups of compounds (reviewedin e.g. Fischer, P. M. Curr. Opin. Drug Discovery Dev. 2001, 4, 623-634)have been found to possess anti-proliferative properties by virtue ofCDK-specific ATP antagonism.

At a molecular level mediation of cdk/cyclin complex activity requires aseries of stimulatory and inhibitory phosphorylation, ordephosphorylation, events. Cdk phosphorylation is performed by a groupof cdk activating kinases (CAKs) and/or kinases such as wee1, Myt1 andMik1. Dephosphorylation is performed by phosphatases such as cdc25(a &c), pp2a, or KAP. Cdk/cyclin complex activity may be further regulatedby two families of endogenous cellular proteinaceous inhibitors: theKip/Cip family, or the INK family. The INK proteins specifically bindcdk4 and cdk6. p16^(ink4) (also known as MTS1) is a potential tumoursuppressor gene that is mutated, or deleted, in a large number ofprimary cancers. The Kip/Cip family contains proteins such asp21^(Cip1,Waf1), p27^(Kip1) and p57^(kip2). As discussed previously p21is induced by p53 and is able to inactivate the cdk2/cyclin(E/A) andcdk4/cyclin(D1/D2/D3) complexes. Atypically low levels of p27 expressionhave been observed in breast, colon and prostate cancers. Converselyover expression of cyclin E in solid tumours has been shown to correlatewith poor patient prognosis. Over expression of cyclin D1 has beenassociated with oesophageal, breast, squamous, and non-small cell lungcarcinomas.

The pivotal roles of cdks, and their associated proteins, inco-ordinating and driving the cell cycle in proliferating cells havebeen outlined above. Some of the biochemical pathways in which cdks playa key role have also been described. The development of monotherapiesfor the treatment of proliferative disorders, such as cancers, usingtherapeutics targeted generically at cdks, or at specific cdks, istherefore potentially highly desirable. Cdk inhibitors could conceivablyalso be used to treat other conditions such as viral infections,autoimmune diseases and neuro-degenerative diseases, amongst others. Cdktargeted therapeutics may also provide clinical benefits in thetreatment of the previously described diseases when used in combinationtherapy with either existing, or new, therapeutic agents. Cdk targetedanticancer therapies could potentially have advantages over many currentantitumour agents as they would not directly interact with DNA andshould therefore reduce the risk of secondary tumour development.

Thus, there is a continued need to find new therapeutic agents to treathuman diseases. Accordingly, there is a great need to develop inhibitorsof protein kinases, such as CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 andCDK9.

SUMMARY OF THE INVENTION

There remains a need for new treatments and therapies for proteinkinase-associated disorders. There is also a need for compounds usefulin the treatment or prevention or amelioration of one or more symptomsof cancer, transplant rejections, and autoimmune diseases. Furthermore,there is a need for methods for modulating the activity of proteinkinases, such as CDK1, CDK2, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9,using the compounds provided herein. In one aspect, the inventionprovides a compound of Formula I:

In an embodiment, the present invention includes a compound of formulaI:

or pharmaceutically acceptable salts, wherein

X is CR⁹, or N;

R¹ is C₁₋₈alkyl, CN, C(O)OR⁴ or CONR⁵R⁶, a 5-14 membered heteroarylgroup, or a 3-14 membered cycloheteroalkyl group;

R² is C₁₋₈alkyl, C₃₋₁₄cycloalkyl, or a 5-14 membered heteroaryl group,and wherein R² may be substituted with one or more C₁₋₈alkyl, or OH;

L is a bond, C₁₋₈alkylene, C(O), or C(O)NR¹⁰, and wherein L may besubstituted or unsubstituted;

Y is H, R¹¹, NR¹²R¹³, OH, or Y is part of the following group

where Y is CR⁹ or N; where 0-3 R⁸ may be present, and R⁸ is C₁₋₈alkyl,oxo, halogen, or two or more R⁸ may form a bridged alkyl group;

W is CR⁹, or N, or O (where W is O, R3 is absent);

R³ is H, C₁₋₈alkyl, C₁₋₈alkylR¹⁴, C₃₋₁₄cycloalkyl, C(O)C₁₋₈ alkyl,C₁₋₈haloalkyl, C₁₋₈alkylOH, C(O)NR¹⁴R¹⁵, C₁₋₈cyanoalkyl, C(O)R¹⁴,C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵, C₀₋₈alkylC(O)OR¹⁴, NR¹⁴R¹⁵, SO₂C₁₋₈alkyl,C₁₋₈alkylC₃₋₁₄cycloalkyl, C(O)C₁₋₈alkylC₃₋₁₄cycloalkyl, C₁₋₈alkoxy, orOH which may be substituted or unsubstituted when R³ is not H.

R⁹ is H or halogen;

R⁴, R⁵, R⁶, R⁷, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, and R¹⁵ are each independentlyselected from H, C₁₋₈alkyl, C₃₋₁₄ cycloalkyl, a 3-14 memberedcycloheteroalkyl group, a C₆₋₁₄ aryl group, a 5-14 membered heteroarylgroup, alkoxy, C(O)H, C(N)OH, C(N)OCH₃, C(O)C₁₋₃alkyl, C₁₋₈alkylNH₂,C₁₋₆alkylOH, and wherein R⁴, R⁵, R⁶, R⁷, R¹⁰, R¹¹, R¹², and R¹³, R¹⁴,and R¹⁵ when not H may be substituted or unsubstituted;

m and n are independently 0-2; and

wherein L, R³, R⁴, R⁵, R⁶, R⁷, R¹⁰, R¹¹, R¹², and R¹³, R¹⁴, and R¹⁵ maybe substituted with one or more of C₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl,C₃₋₁₄cycloalkyl, 5-14 membered heteroaryl group, C₆₋₁₄aryl group, a 3-14membered cycloheteroalkyl group, OH, (O), CN, alkoxy, halogen, or NH₂.

DETAILED DESCRIPTION OF THE INVENTION

In an embodiment, the present invention includes compounds of formula Iwherein Y is H, OH, or Y is part of the following group

where Y is N and W is CR⁹, or N; and where 0-2 R⁸ may be present, and R⁸is C₁₋₈alkyl, oxo, or two or more R⁸ may form a bridged alkyl group. Inan embodiment, Y is N and W is N. In an embodiment, m is 1 or 2. Inanother embodiment, n is 1 or 2. In an embodiment, m is 1 and n is 2. Inanother embodiment, m is 2 and n is 1. In a further embodiment, both mand n are 1.

In an embodiment, there are 0-2 R⁸ present in compounds of formula (I).It is understood that when there are zero les, that H is attached to thecarbons of the cyclic structure.

In an embodiment, R⁸ is methyl, ethyl, propyl, butyl, oxo, or two R⁸ canform a bridged (cycloalkyl) group, such as cyclobutyl, cyclopentyl, orcyclohexyl. In an embodiment, R⁸ is methyl. In another embodiment no R⁸is present.

In an embodiment, the present invention includes compounds of formula Iwherein R³ is H, C₁₋₈alkyl, such as methyl, ethyl, propyl, isopropyl,butyl, pentyl, or hexyl; C₃₋₁₄cycloalkyl, such as cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl; C(O)C₁₋₈ alkyl, such as C(O)CH₃,C(O)CH₂CH₃, or C(O)CH₂CH₂CH₃; C₁₋₈alkylOH, such as CH₂OH, CH₂CH₂OH,CHOHCH₃, CH₂CH₂CH₂OH, CHOHCH₂CH₃, or CH₂CHOHCH₃; C₁₋₈cyanoalkyl, such asCH₂CN, or CH₂CH₂CN; C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵, such asCH₂C(O)CH₂NR¹⁴R¹⁵; C₀₋₈alkylC(O)OR¹⁴, NR¹⁴R¹⁵, C₁₋₈alkylC₃₋₁₄cycloalkyl,C(O)C₁₋₈alkylC₃₋₁₄cycloalkyl, C₀₋₈alkoxy, C₁₋₈alkylR¹⁴, C₁₋₈haloalkyl,or C(O)R¹⁴, which may be substituted with one or more of OH, CN, F, orNH₂, and wherein R¹⁴ and R¹⁵ are each independently selected from H,C₁₋₈alkyl, C₃₋₁₄cycloalkyl, alkoxy, C(O)C₁₋₃alkyl, C₁₋₈alkylNH₂, orC₁₋₆alkylOH.

In an embodiment, R¹⁴, and R¹⁵ are each independently selected from H,C₁₋₈alkyl, such as methyl, ethyl, propyl, butyl, pentyl, or hexyl;C₃₋₁₄cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl; a 3-14 membered cycloheteroalkyl group, such as morpoholine,piperidine, or piperazine; a C₆₋₁₄ aryl group, such as phenyl; a 5-14membered heteroaryl group, such as pyridine, pyrimidine, or pyridazine;alkoxy, such as methoxy, ethoxy, or propoxy; C(O)H, C(N)OH, C(N)OCH₃,C(O)C₁₋₃alkyl, such as C(O)CH₃, C(O)CH₂CH₃, or C(O)CH₂CH₂CH₃;C₁₋₈alkylNH₂, such as methyleneNH₂, ethyleneNH₂, or propyleneNH₂; C₁₋₆alkylOH, such as methyleneOH, ethyleneOH, or propyleneOH; and R14 andR15 when not H may be unsubstituted or substituted with one or more ofC₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl, C₃₋₁₄cycloalkyl, 5-14 memberedheteroaryl group, C₆₋₁₄aryl group, a 3-14 membered cycloheteroalkylgroup, OH, (O), CN, alkoxy, halogen, or NH₂.

In another embodiment, the present invention includes compound offormula I wherein R³ is H, C₁₋₈alkyl, such as methyl, ethyl, propyl, orisopropyl; or C₁₋₈alkylOH, such as CH₂OH, or CH₂CH₂OH. In anotherembodiment, R3 is H, isopropyl, CH₂OH, or CH₂CH₂OH. In anotherembodiment, R³ is H.

In another embodiment, the present invention includes compounds offormula I wherein L is a bond, C₁₋₈alkylene, such as —CH₂—, —CH₂CH₂—, or—CH₂CH₂CH₂—; C(O)NH, or C(O).

In another embodiment, the present invention includes compounds offormula I wherein R² is C₃₋₁₄cycloalkyl; such as cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl.

In another embodiment, the present invention includes compounds offormula I wherein R² is cyclopentyl.

In another embodiment, the present invention includes compounds offormula I wherein R¹ is CN, C(O)OR⁴, CONR⁵R⁶, or a 5-14 memberedheteroaryl group.

In another embodiment, the present invention includes compounds offormula I wherein R¹ is CONR⁵R⁶, and R⁵ and R⁶ are C₁₋₈alkyl. In anotherembodiment, le is CONR⁵R⁶ where R⁵ and R⁶ are methyl. In anotherembodiment, le is CN.

In another embodiment, the present invention includes compounds offormula I wherein X is CR⁹, and R⁹ is H or halogen, such as Cl, F, Br,or I.

In another embodiment, the present invention includes compounds offormula I wherein one X is N and the other X is CR⁹. In an embodiment,the present invention includes compounds of formula (I), such as:

In another embodiment, the present invention includes compounds offormula I wherein X is CR⁹ and Y is

where m and n are 1, and Y and W are N.

In one embodiment the compound is a compound of formula I wherein L is abond, C₁₋₈alkylene, or C(O)NH, or C(O); and Y is H, OH, or Y is part ofthe following group

where Y is N and W is CR⁹, or N; where 0-2 R⁸ may be present, andR⁸ is C₁₋₈alkyl, oxo, or two or more R⁸ may link to form a bridged alkylgroup andR³ is H, C₁₋₈alkyl, C₁₋₈alkylR¹⁴, C₁₋₈haloalkyl, C(O)C₁₋₈ alkyl,C₀₋₈alkylOH, C(O)R¹⁴, or C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵,C₀₋₈alkylC(O)OR¹⁴, or NR¹⁴R¹⁵; andR¹⁴ and R¹⁵ are each independently selected from H, C₁₋₈alkyl, C₃₋₁₄cycloalkyl, alkoxy, C(O)C₁₋₃alkyl, C₁₋₈alkylNH₂, C₁₋₆ alkylOH.

In one embodiment the compound is a compound of formula I wherein R³ isH, C₁₋₈alkyl, C₃₋₁₄cycloalkyl, C(O)C₁₋₈ alkyl, C₀₋₈alkylOH,C₁₋₈cyanoalkyl, C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵, C₀₋₈alkylC(O)OR¹⁴,NR¹⁴R¹⁵, C₁₋₈alkylC₃₋₁₄cycloalkyl, C(O)C₁₋₈alkylC₃₋₁₄cycloalkyl,C₀₋₈alkoxy, which may be substituted with one or more of OH, CN, F, orNH₂.

In one embodiment the compound is a compound of formula I wherein R³ isH or C₁₋₈alkyl.

In one embodiment the compound is a compound of formula I wherein R¹ isC(O)OR⁴, CONR⁵R⁶, or a 5-14 membered heteroaryl group.

In one embodiment, the present invention includes compounds of formula Iwherein Y is

where m and n are 1 or 2, and Y and W are N.

In one embodiment the compound is a compound of formula I wherein L is abond.

In one embodiment the compound is a compound of formula I wherein L is abond Y is not H.

In another embodiment, the present invention includes compounds offormula I(a):

and a pharmaceutically acceptable salt thereof, wherein:R⁵⁰ is CONR⁵⁴R⁵⁵, or CN;R⁵¹ is C₃₋₁₄cycloalkyl which may be unsubstituted or substituted byC₁₋₃alkyl, or OH;

Z is CH or N; and

V is NR⁵⁶ or CHR⁵⁷;R⁵⁴ and R⁵⁵ are independently H, C₁₋₃alkyl,R⁵², R⁵³, R⁵⁶, and R⁵⁷ are independently H, C₁₋₈alkyl, C₃₋₁₄cycloalkyl,C₁₋₈haloalkyl, NR⁵⁸R⁵⁹, C(O)OR⁶⁰, C(O)C₁₋₈alkyl,C₀₋₈alkylC(O)C₀₋₈alkyl-NR⁶¹R⁶², C₁₋₈alkoxy, C₁₋₈alkylOR⁶³,C(O)-5-14cycloheteroalkyl group, C₃₋₁₄cycloalkyl group, each of whichwhen not H may be substituted by one or more of C₁₋₈alky, OH, or CN;R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², and R⁶³ are H or C₁₋₈alkyl.

In an embodiment of the present invention, formula I(a) includescompounds where R⁵⁰ is CONR⁵³R⁵⁵, and R⁵⁴ and R⁵⁵ are H, methyl, orethyl. In another embodiment, R⁵⁴ and R⁵⁵ are both methyl.

In another embodiment, formula I(a) includes compounds where R⁵¹ iscyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. In anotherembodiment, the invention includes compounds where R⁵¹ is cyclopentyl.

In another embodiment, formula I(a) includes compounds where Z is N. Inanother embodiment, the present invention includes compounds where V isNR⁵⁶. In another embodiment, the present invention includes compoundswhere V is NR⁵⁶, and R⁵⁶ is H, methyl, ethyl, propyl which may besubstituted by OH. In another embodiment, R⁵⁶ is isopropyl. In anotherembodiment, R⁵⁶ is H. In yet another embodiment, R⁵⁶ is —CH₂CH₂OH.

In another embodiment, the present invention includes a method oftreating a disease, disorder or syndrome associated with CDK 4inhibition, said method comprising administering a compound according toformula I or I(a) or its prodrug or pharmaceutical compositioncomprising the compound of formula I or I(a) or its prodrug andpharmaceutically acceptable excipients to a subject in need thereof.

In another embodiment, the present invention includes a method oftreating a disease associated with CDK 4 inhibition, wherein thedisease, disorder or syndrome is hyperproliferative in a subject,wherein subject is an animal including humans, selected from a groupcomprising cancer and inflammation.

In another embodiment, the present invention includes a method ofinhibiting a cyclin dependent kinase (e.g. cdk-4), which methodcomprises contacting the kinase with a kinase-inhibiting compoundaccording to formula I or I(a).

In another embodiment, the present invention includes a method ofmodulating a cellular process (for example cell division) by inhibitingthe activity of a cyclin dependent kinase using a compound according toformula I or I(a).

In another embodiment, the present invention includes a compound offormula I or I(a) for use in the prophylaxis or treatment of a diseasestate as described herein.

In another embodiment, the present invention includes the use of acompound of formula I or I(a) for the manufacture of a medicament,wherein the medicament is for any one or more of the uses definedherein.

In another embodiment, the present invention includes a pharmaceuticalcomposition comprising a compound of formula I or I(a) and apharmaceutically acceptable carrier.

In another embodiment, the present invention includes a pharmaceuticalcomposition comprising a compound of formula I or I(a) and apharmaceutically acceptable carrier in a form suitable for oraladministration.

The phrase “pharmaceutically acceptable” refers to molecular entitiesand compositions that are physiologically tolerable and do not typicallyproduce an allergic or similar untoward reaction, such as gastric upset,dizziness and the like, when administered to a human. Preferably, asused herein, the term “pharmaceutically acceptable” means approved by aregulatory agency of the Federal or a state government or listed in theU.S. Pharmacopeia or other generally recognized pharmacopeia for use inanimals, and more particularly in humans.

The term “carrier” refers to a diluent, adjuvant, excipient, or vehiclewith which the compound is administered. Such pharmaceutical carrierscan be sterile liquids, such as water and oils, including those ofpetroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. Water or aqueoussolution saline solutions and aqueous dextrose and glycerol solutionsare preferably employed as carriers, particularly for injectablesolutions. Suitable pharmaceutical carriers are described in“Remington's Pharmaceutical Sciences” by E. W. Martin.

The phrase “therapeutically effective amount” is used herein to mean anamount sufficient to reduce by at least about 15 percent, preferably byat least 50 percent, more preferably by at least 90 percent, and mostpreferably prevent, a clinically significant deficit in the activity,function and response of the host. Alternatively, a therapeuticallyeffective amount is sufficient to cause an improvement in a clinicallysignificant condition/symptom in the host.

“Agent” refers to all materials that may be used to preparepharmaceutical and diagnostic compositions, or that may be compounds,nucleic acids, polypeptides, fragments, isoforms, variants, or othermaterials that may be used independently for such purposes, all inaccordance with the present invention.

“Analog” as used herein, refers to a small organic compound, anucleotide, a protein, or a polypeptide that possesses similar oridentical activity or function(s) as the compound, nucleotide, proteinor polypeptide or compound having the desired activity and therapeuticeffect of the present invention. (e.g., inhibition of tumor growth), butneed not necessarily comprise a sequence or structure that is similar oridentical to the sequence or structure of the preferred embodiment

“Derivative” refers to either a compound, a protein or polypeptide thatcomprises an amino acid sequence of a parent protein or polypeptide thathas been altered by the introduction of amino acid residuesubstitutions, deletions or additions, or a nucleic acid or nucleotidethat has been modified by either introduction of nucleotidesubstitutions or deletions, additions or mutations. The derivativenucleic acid, nucleotide, protein or polypeptide possesses a similar oridentical function as the parent polypeptide.

As used herein, “halo” or “halogen” refers to fluoro, chloro, bromo, andiodo.

As used herein, “alkyl” refers to a straight-chain or branched saturatedhydrocarbon group. In some embodiments, an alkyl group can have from 1to 10 carbon atoms (e.g., from 1 to 8 carbon atoms). Examples of alkylgroups include methyl (Me), ethyl (Et), propyl (e.g., n-propyl andisopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl, t-butyl), pentylgroups (e.g., n-pentyl, isopentyl, neopentyl), hexyl (e.g., n-hexyl andits isomers), and the like. A lower alkyl group typically has up to 4carbon atoms. Examples of lower alkyl groups include methyl, ethyl,propyl (e.g., n-propyl and isopropyl), and butyl groups (e.g., n-butyl,isobutyl, s-butyl, t-butyl). In an embodiment an alkyl group, or two ormore alkyl groups may form a bridged alkyl group. This is where an alkylgroup links across another group (particularly shown in cyclic groups),forming a ring bridged by an alkyl chain, i.e., forming a bridged fusedring. This is shown, but not limited to where two or more R⁸ groups fora bridged alkyl group across the Y ring group forming a ring bridged byan alkyl chain.

As used herein, “alkenyl” refers to a straight-chain or branched alkylgroup having one or more carbon-carbon double bonds. In someembodiments, an alkenyl group can have from 2 to 10 carbon atoms (e.g.,from 2 to 8 carbon atoms). Examples of alkenyl groups include ethenyl,propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl,hexadienyl groups, and the like. The one or more carbon-carbon doublebonds can be internal (such as in 2-butene) or terminal (such as in1-butene).

As used herein, “alkynyl” refers to a straight-chain or branched alkylgroup having one or more carbon-carbon triple bonds. In someembodiments, an alkynyl group can have from 2 to 10 carbon atoms (e.g.,from 2 to 8 carbon atoms). Examples of alkynyl groups include ethynyl,propynyl, butynyl, pentynyl, and the like. The one or more carbon-carbontriple bonds can be internal (such as in 2-butyne) or terminal (such asin 1-butyne).

As used herein, “alkoxy” refers to an —O— alkyl group. Examples ofalkoxy groups include methoxy, ethoxy, propoxy (e.g., n-propoxy andisopropoxy), t-butoxy groups, and the like. As used herein, “alkylthio”refers to an —S— alkyl group. Examples of alkylthio groups includemethylthio, ethylthio, propylthio (e.g., n-propylthio andisopropylthio), t-butylthio groups, and the like.

The term “carbalkoxy” refers to an alkoxycarbonyl group, where theattachment to the main chain is through the carbonyl group (C(O)).Examples include but are not limited to methoxy carbonyl, ethoxycarbonyl, and the like.

As used herein, “oxo” refers to a double-bonded oxygen (i.e., ═O). It isalso to be understood that the terminology C(O) refers to a —C═O group,whether it be ketone, aldehyde or acid or acid derivative. Similarly,S(O) refers to a —S═O group.

As used herein, “haloalkyl” refers to an alkyl group having one or morehalogen substituents. In some embodiments, a haloalkyl group can have 1to 10 carbon atoms (e.g., from 1 to 8 carbon atoms). Examples ofhaloalkyl groups include CF₃, C₂F₅, CHF₂, CH₂F, CCl₃, CHCl₂, CH₂Cl,C₂Cl₅, and the like. Perhaloalkyl groups, i.e., alkyl groups wherein allof the hydrogen atoms are replaced with halogen atoms (e.g., CF₃ andC₂F₅), are included within the definition of “haloalkyl.” For example, aC₁₋₁₀ haloalkyl group can have the formula —C_(i)H_(2i+1−j)X_(i),wherein X is F, Cl, Br, or I, i is an integer in the range of 1 to 10,and j is an integer in the range of 0 to 21, provided that j is lessthan or equal to 2i+1.

As used herein, “cycloalkyl” refers to a non-aromatic carbocyclic groupincluding cyclized alkyl, alkenyl, and alkynyl groups. A cycloalkylgroup can be monocyclic (e.g., cyclohexyl) or polycyclic (e.g.,containing fused, bridged, and/or spiro ring systems), wherein thecarbon atoms are located inside or outside of the ring system. Acycloalkyl group, as a whole, can have from 3 to 14 ring atoms (e.g.,from 3 to 8 carbon atoms for a monocyclic cycloalkyl group and from 7 to14 carbon atoms for a polycyclic cycloalkyl group). Any suitable ringposition of the cycloalkyl group can be covalently linked to the definedchemical structure. Examples of cycloalkyl groups include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl,cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinyl,norcaryl, adamantyl, and spiro[4.5]decanyl groups, as well as theirhomologs, isomers, and the like.

As used herein, “heteroatom” refers to an atom of any element other thancarbon or hydrogen and includes, for example, nitrogen, oxygen, sulfur,phosphorus, and selenium.

As used herein, “cycloheteroalkyl” refers to a non-aromatic cycloalkylgroup that contains at least one (e.g., one, two, three, four, or five)ring heteroatom selected from O, N, and S, and optionally contains oneor more (e.g., one, two, or three) double or triple bonds. Acycloheteroalkyl group, as a whole, can have from 3 to 14 ring atoms andcontains from 1 to 5 ring heteroatoms (e.g., from 3-6 ring atoms for amonocyclic cycloheteroalkyl group and from 7 to 14 ring atoms for apolycyclic cycloheteroalkyl group). The cycloheteroalkyl group can becovalently attached to the defined chemical structure at anyheteroatom(s) or carbon atom(s) that results in a stable structure. Oneor more N or S atoms in a cycloheteroalkyl ring may be oxidized (e.g.,morpholine N-oxide, thiomorpholine S-oxide, thiomorpholine S,S-dioxide).Cycloheteroalkyl groups can also contain one or more oxo groups, such asphthalimidyl, piperidonyl, oxazolidinonyl, 2,4(1H,3H)-dioxo-pyrimidinyl,pyridin-2(1H)-onyl, and the like. Examples of cycloheteroalkyl groupsinclude, among others, morpholinyl, thiomorpholinyl, pyranyl,imidazolidinyl, imidazolinyl, oxazolidinyl, pyrazolidinyl, pyrazolinyl,pyrrolidinyl, pyrrolinyl, tetrahydrofuranyl, tetrahydrothienyl,piperidinyl, piperazinyl, azetidine, and the like.

As used herein, “aryl” refers to an aromatic monocyclic hydrocarbon ringsystem or a polycyclic ring system where at least one of the rings inthe ring system is an aromatic hydrocarbon ring and any other aromaticrings in the ring system include only hydrocarbons. In some embodiments,a monocyclic aryl group can have from 6 to 14 carbon atoms and apolycyclic aryl group can have from 8 to 14 carbon atoms. The aryl groupcan be covalently attached to the defined chemical structure at anycarbon atom(s) that result in a stable structure. In some embodiments,an aryl group can have only aromatic carbocyclic rings, e.g., phenyl,1-naphthyl, 2-naphthyl, anthracenyl, phenanthrenyl groups, and the like.In other embodiments, an aryl group can be a polycyclic ring system inwhich at least one aromatic carbocyclic ring is fused (i.e., having abond in common with) to one or more cycloalkyl or cycloheteroalkylrings. Examples of such aryl groups include, among others, benzoderivatives of cyclopentane (i.e., an indanyl group, which is a5,6-bicyclic cycloalkyl/aromatic ring system), cyclohexane (i.e., atetrahydronaphthyl group, which is a 6,6-bicyclic cycloalkyl/aromaticring system), imidazoline (i.e., a benzimidazolinyl group, which is a5,6-bicyclic cycloheteroalkyl/aromatic ring system), and pyran (i.e., achromenyl group, which is a 6,6-bicyclic cycloheteroalkyl/aromatic ringsystem). Other examples of aryl groups include benzodioxanyl,benzodioxolyl, chromanyl, indolinyl groups, and the like.

As used herein, “heteroaryl” refers to an aromatic monocyclic ringsystem containing at least one ring heteroatom selected from O, N, and Sor a polycyclic ring system where at least one of the rings in the ringsystem is aromatic and contains at least one ring heteroatom. Aheteroaryl group, as a whole, can have from 5 to 14 ring atoms andcontain 1-5 ring heteroatoms. In some embodiments, heteroaryl groups caninclude monocyclic heteroaryl rings fused to one or more aromaticcarbocyclic rings, non-aromatic carbocyclic rings, or non-aromaticcycloheteroalkyl rings. The heteroaryl group can be covalently attachedto the defined chemical structure at any heteroatom or carbon atom thatresults in a stable structure. Generally, heteroaryl rings do notcontain O—O, S—S, or S—O bonds. However, one or more N or S atoms in aheteroaryl group can be oxidized (e.g., pyridine N-oxide, thiopheneS-oxide, thiophene S,S-dioxide). Examples of such heteroaryl ringsinclude pyrrolyl, furyl, thienyl, pyridyl, pyrimidyl, pyridazinyl,pyrazinyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, isothiazolyl,thiazolyl, thiadiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, indolyl,isoindolyl, benzofuryl, benzothienyl, quinolyl, 2-methylquinolyl,isoquinolyl, quinoxalyl, quinazolyl, benzotriazolyl, benzimidazolyl,benzothiazolyl, benzisothiazolyl, benzisoxazolyl, benzoxadiazolyl,benzoxazolyl, cinnolinyl, 1H-indazolyl, 2H-indazolyl, indolizinyl,isobenzofuyl, naphthyridinyl, phthalazinyl, pteridinyl, purinyl,oxazolopyridinyl, thiazolopyridinyl, imidazopyridinyl, furopyridinyl,thienopyridinyl, pyridopyrimidinyl, pyridopyrazinyl, pyridopyridazinyl,thienothiazolyl, thienoxazolyl, thienoimidazolyl groups, and the like.Further examples of heteroaryl groups include 4,5,6,7-tetrahydroindolyl,tetrahydroquinolinyl, benzothienopyridinyl, benzofuropyridinyl groups,and the like.

The term “lower alkenyl” refers to a alkenyl group which contains 2-6carbon atoms. An alkenyl group is a hydrocarbyl group containing atleast one carbon-carbon double bond. As defined herein, it may beunsubstituted or substituted with the substituents described herein. Thecarbon-carbon double bonds may be between any two carbon atoms of thealkenyl group. It is preferred that it contains 1 or 2 carbon-carbondouble bonds and more preferably one carbon-carbon double bond. Thealkenyl group may be straight chained or branched. Examples include butare not limited to ethenyl, 1-propenyl, 2-propenyl, 1-butenyl,2-butenyl, 2-methyl-1-propenyl, 1, 3-butadienyl, and the like.

The term “lower alkynyl”, as used herein, refers to an alkynyl groupcontaining 2-6 carbon atoms. An alkynyl group is a hydrocarbyl groupcontaining at least one carbon-carbon triple bond. The carbon-carbontriple bond may be between any two carbon atom of the alkynyl group. Inan embodiment, the alkynyl group contains 1 or 2 carbon-carbon triplebonds and more preferably one carbon-carbon triple bond. The alkynylgroup may be straight chained or branched. Examples include but are notlimited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl and thelike.

The present invention includes all pharmaceutically acceptableisotopically-labeled compounds of the invention, i.e. compounds offormula (I) or I(a), wherein one or more atoms are replaced by atomshaving the same atomic number, but an atomic mass or mass numberdifferent from the atomic mass or mass number usually found in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention comprises isotopes of hydrogen, such as ²H and ³H, carbon,such as ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F,iodine, such as ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen,such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, and sulphur, such as³⁵S.

Certain isotopically-labelled compounds of formula (I) or I(a), forexample, those incorporating a radioactive isotope, are useful in drugand/or substrate tissue distribution studies. The radioactive isotopestritium, i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful forthis purpose in view of their ease of incorporation and ready means ofdetection.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N can be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy.

Isotopically-labeled compounds of formula (I) or I(a) can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described in the accompanying Examplesand Preparations using an appropriate isotopically-labeled reagents inplace of the non-labeled reagent previously employed.

Biological Activity

The compounds of the formulae (I) or I(a) and sub-groups thereof areinhibitors of cyclin dependent kinases. For example, compounds of theinvention are inhibitors of cyclin dependent kinases, and in particularcyclin dependent kinases selected from CDK1, CDK2, CDK3, CDK4, CDK5,CDK6 and CDK9, and more particularly selected from CDK1, CDK2, CDK3,CDK4, CDK5 and CDK9.

Compounds of the invention also have activity against glycogen synthasekinase-3 (GSK-3). As a consequence of their activity in modulating orinhibiting CDK and glycogen synthase kinase, they are expected to beuseful in providing a means of arresting, or recovering control of, thecell cycle in abnormally dividing cells. It is therefore anticipatedthat the compounds will prove useful in treating or preventingproliferative disorders such as cancers. It is also envisaged that thecompounds of the invention will be useful in treating conditions such asviral infections, type II or non-insulin dependent diabetes mellitus,autoimmune diseases, head trauma, stroke, epilepsy, neurodegenerativediseases such as Alzheimer's, motor neurone disease, progressivesupranuclear palsy, corticobasal degeneration and Pick's disease forexample autoimmune diseases and neurodegenerative diseases.

One sub-group of disease states and conditions where it is envisagedthat the compounds of the invention will be useful consists of viralinfections, autoimmune diseases and neurodegenerative diseases.

CDKs play a role in the regulation of the cell cycle, apoptosis,transcription, differentiation and CNS function. Therefore, CDKinhibitors could be useful in the treatment of diseases in which thereis a disorder of proliferation, apoptosis or differentiation such ascancer. In particular RB+ve tumours may be particularly sensitive to CDKinhibitors. These include tumours harbouring mutations in ras, Raf,Growth Factor Receptors or over-expression of Growth Factor Receptors.Furthermore tumours with hypermethylated promoter regions of CDKinhibitors as well as tumours over-expressing cyclin partners of thecyclin dependent kinases may also display sensitivity. RB-ve tumours mayalso be sensitive to CDK inhibitors.

Examples of cancers which may be inhibited include, but are not limitedto, a carcinoma, for example a carcinoma of the bladder, breast, colon(e.g. colorectal carcinomas such as colon adenocarcinoma and colonadenoma), kidney, epidermis, liver, lung, for example adenocarcinoma,small cell lung cancer and non-small cell lung carcinomas, oesophagus,gall bladder, ovary, pancreas e.g. exocrine pancreatic carcinoma,stomach, cervix, thyroid, nose, head and neck, prostate, or skin, forexample squamous cell carcinoma; a hematopoietic tumour of lymphoidlineage, for example leukemia, acute lymphocytic leukemia, chroniclymphocytic leukaemia, B-cell lymphoma (such as diffuse large B celllymphoma), T-cell lymphoma, multiple myeloma, Hodgkin's lymphoma,non-Hodgkin's lymphoma, hairy cell lymphoma, or Burkett's lymphoma; ahematopoietic tumour of myeloid lineage, for example acute and chronicmyelogenous leukemias, myelodysplastic syndrome, or promyelocyticleukemia; thyroid follicular cancer; a tumour of mesenchymal origin, forexample fibrosarcoma or habdomyosarcoma; a tumour of the central orperipheral nervous system, for example astrocytoma, neuroblastoma,glioma or schwannoma; melanoma; seminoma; teratocarcinoma; osteosarcoma;xeroderma pigmentosum; keratoctanthoma; thyroid follicular cancer; orKaposi's sarcoma.

The cancers may be cancers which are sensitive to inhibition of any oneor more cyclin dependent kinases selected from CDK1, CDK2, CDK3, CDK4,CDK5 and CDK6, for example, one or more CDK kinases selected from CDK1,CDK2, CDK4 and CDK5, e.g. CDK1 and/or CDK2. Whether or not a particularcancer is one which is sensitive to inhibition by a cyclin dependentkinase inhibitor may be determined by means of a cell growth assay asset out in the examples below or by a method as set out in the sectionheaded “Methods of Diagnosis”.

CDKs are also known to play a role in apoptosis, proliferation,differentiation and transcription and therefore CDK inhibitors couldalso be useful in the treatment of the following diseases other thancancer; viral infections, for example herpes virus, pox virus,Epstein-Barr virus, Sindbis virus, adenovirus, HIV, HPV, HCV and HCMV;prevention of AIDS development in HIV-infected individuals; chronicinflammatory diseases, for example systemic lupus erythematosus,autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis,inflammatory bowel disease, and autoimmune diabetes mellitus;cardiovascular diseases for example cardiac hypertrophy, restenosis,atherosclerosis; neurodegenerative disorders, for example Alzheimer'sdisease, AIDS-related dementia, Parkinson's disease, amyotropic lateralsclerosis, retinitis pigmentosa, spinal muscular atropy and cerebellardegeneration; glomerulonephritis; myelodysplastic syndromes, ischemicinjury associated myocardial infarctions, stroke and reperfusion injury,arrhythmia, atherosclerosis, toxin-induced or alcohol related liverdiseases, haematological diseases, for example, chronic anemia andaplastic anemia; degenerative diseases of the musculoskeletal system,for example, osteoporosis and arthritis, aspirin-sensitiverhinosinusitis, cystic fibrosis, multiple sclerosis, kidney diseases,ophthalmic diseases including age related macular degeneration, uveitis,and cancer pain.

It has also been discovered that some cyclin-dependent kinase inhibitorscan be used in combination with other anticancer agents. For example,the cyclin-dependent kinase inhibitor flavopiridol has been used withother anticancer agents in combination therapy.

Thus, in the pharmaceutical compositions, uses or methods of thisinvention for treating a disease or condition comprising abnormal cellgrowth, the disease or condition comprising abnormal cell growth in oneembodiment is a cancer.

One group of cancers includes human breast cancers (e.g. primary breasttumours, node-negative breast cancer, invasive duct adenocarcinomas ofthe breast, non-endometrioid breast cancers); and mantle cell lymphomas.In addition, other cancers are colorectal and endometrial cancers.

Another sub-set of cancers includes hematopoietic tumours of lymphoidlineage, for example leukemia, chronic lymphocytic leukaemia, mantlecell lymphoma and B-cell lymphoma (such as diffuse large B celllymphoma).

One particular cancer is chronic lymphocytic leukaemia.

Another particular cancer is mantle cell lymphoma.

Another particular cancer is diffuse large B cell lymphoma

Another sub-set of cancers includes breast cancer, ovarian cancer, coloncancer, prostate cancer, oesophageal cancer, squamous cancer andnon-small cell lung carcinomas.

Another sub-set of cancers includes breast cancer, pancreatic cancer,colorectal cancer, lung cancer, and melanoma.

A further sub-set of cancers, namely cancers wherein compounds havingCDK4 inhibitory activity may be of particular therapeutic benefit,comprises retinoblastomas, small cell lung carcinomas, non-small lungcarcinomas, sarcomas, gliomas, pancreatic cancers, head, neck and breastcancers and mantle cell lymphomas.

Another sub-set of cancers wherein compounds having CDK4 inhibitoryactivity may be of particular therapeutic benefit comprises small celllung cancer, non-small cell lung cancer, pancreatic cancer, breastcancer, glioblastoma multiforme, T cell ALL and mantle cell lymphoma. Afurther subset of cancers which the compounds of the invention may beuseful in the treatment of includes sarcomas, leukemias, glioma,familial melanoma and melanoma.

Methods of Diagnosis

Prior to administration of a compound of the formula (I) or I(a), apatient may be screened to determine whether a disease or condition fromwhich the patient is or may be suffering is one which would besusceptible to treatment with a compound having activity against cyclindependent kinases. For example, a biological sample taken from a patientmay be analysed to determine whether a condition or disease, such ascancer, that the patient is or may be suffering from is one which ischaracterised by a genetic abnormality or abnormal protein expressionwhich leads to over-activation of CDKs or to sensitisation of a pathwayto normal CDK activity. Examples of such abnormalities that result inactivation or sensitisation of the CDK2 signal include up-regulation ofcyclin E, (Harwell R M, Mull B B, Porter D C, Keyomarsi K.; J Biol Chem.2004 Mar. 26; 279(13):12695-705) or loss of p21 or p27, or presence ofCDCl₄ variants (Raj agopalan H, Jallepalli P V, Rago C, Velculescu V E,Kinzler K W, Vogelstein B, Lengauer C.; Nature. 2004 Mar. 4;428(6978):77-81). Tumours with mutants of CDCl₄ or up-regulation, inparticular over-expression, of cyclin E or loss of p21 or p27 may beparticularly sensitive to CDK inhibitors. The term up-regulationincludes elevated expression or over-expression, including geneamplification (i.e. multiple gene copies) and increased expression by atranscriptional effect, and hyperactivity and activation, includingactivation by mutations.

Thus, the patient may be subjected to a diagnostic test to detect amarker characteristic of up-regulation of cyclin E, or loss of p21 orp27, or presence of CDCl₄ variants. The term diagnosis includesscreening. By marker we include genetic markers including, for example,the measurement of DNA composition to identify mutations of CDCl₄. Theterm marker also includes markers which are characteristic of upregulation of cyclin E, including enzyme activity, enzyme levels, enzymestate (e.g. phosphorylated or not) and mRNA levels of the aforementionedproteins. Tumours with upregulation of cyclin E, or loss of p21 or p27may be particularly sensitive to CDK inhibitors. Tumours maypreferentially be screened for upregulation of cyclin E, or loss of p21or p27 prior to treatment. Thus, the patient may be subjected to adiagnostic test to detect a marker characteristic of up-regulation ofcyclin E, or loss of p21 or p27.

The diagnostic tests are typically conducted on a biological sampleselected from tumour biopsy samples, blood samples (isolation andenrichment of shed tumour cells), stool biopsies, sputum, chromosomeanalysis, pleural fluid, peritoneal fluid, or urine.

It has been found, Raj agopalan et al (Nature. 2004 Mar. 4;428(6978):77-81), that there were mutations present in CDCl₄ (also knownas Fbw7 or Archipelago) in human colorectal cancers and endometrialcancers (Spruck et al, Cancer Res. 2002 Aug. 15; 62(16):4535-9).Identification of individual carrying a mutation in CDCl₄ may mean thatthe patient would be particularly suitable for treatment with a CDKinhibitor. Tumours may preferentially be screened for presence of aCDCl₄ variant prior to treatment. The screening process will typicallyinvolve direct sequencing, oligonucleotide microarray analysis, or amutant specific antibody.

Methods of identification and analysis of mutations and up-regulation ofproteins are well known to a person skilled in the art. Screeningmethods could include, but are not limited to, standard methods such asreverse-transcriptase polymerase chain reaction (RT-PCR) or in-situhybridisation.

In screening by RT-PCR, the level of mRNA in the tumour is assessed bycreating a cDNA copy of the mRNA followed by amplification of the cDNAby PCR. Methods of PCR amplification, the selection of primers, andconditions for amplification, are known to a person skilled in the art.Nucleic acid manipulations and PCR are carried out by standard methods,as described for example in Ausubel, F. M. et al., eds. CurrentProtocols in Molecular Biology, 2004, John Wiley & Sons Inc., or Innis,M. A. et-al., eds. PCR Protocols: a guide to methods and applications,1990, Academic Press, San Diego. Reactions and manipulations involvingnucleic acid techniques are also described in Sambrook et al., 2001,3^(rd) Ed, Molecular Cloning: A Laboratory Manual, Cold Spring HarborLaboratory Press. Alternatively a commercially available kit for RT-PCR(for example Roche Molecular Biochemicals) may be used, or methodologyas set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531;5,192,659, 5,272,057, 5,882,864, and 6,218,529 and incorporated hereinby reference.

An example of an in-situ hybridisation technique for assessing mRNAexpression would be fluorescence in-situ hybridisation (FISH) (seeAngerer, 1987 Meth. Enzymol., 152: 649). Generally, in situhybridization comprises the following major steps: (1) fixation oftissue to be analyzed; (2) prehybridization treatment of the sample toincrease accessibility of target nucleic acid, and to reduce nonspecificbinding; (3) hybridization of the mixture of nucleic acids to thenucleic acid in the biological structure or tissue; (4)post-hybridization washes to remove nucleic acid fragments not bound inthe hybridization, and (5) detection of the hybridized nucleic acidfragments. The probes used in such applications are typically labeled,for example, with radioisotopes or fluorescent reporters. Preferredprobes are sufficiently long, for example, from about 50, 100, or 200nucleotides to about 1000 or more nucleotides, to enable specifichybridization with the target nucleic acid(s) under stringentconditions. Standard methods for carrying out FISH are described inAusubel, F. M. et al., eds. Current Protocols in Molecular Biology,2004, John Wiley & Sons Inc and Fluorescence In Situ Hybridization:Technical Overview by John M. S. Bartlett in Molecular Diagnosis ofCancer, Methods and Protocols, 2nd ed.; ISBN: 1-59259-760-2; March 2004,pps. 077-088; Series: Methods in Molecular Medicine.

Alternatively, the protein products expressed from the mRNAs may beassayed by immunohistochemistry of tumour samples, solid phaseimmunoassay with microtiter plates, Western blotting, 2-dimensionalSDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry and othermethods known in the art for detection of specific proteins. Detectionmethods would include the use of site specific antibodies. The skilledperson will recognize that all such well-known techniques for detectionof upregulation of cyclin E, or loss of p21 or p27, or detection ofCDCl₄ variants could be applicable in the present case.

Therefore, all of these techniques could also be used to identifytumours particularly suitable for treatment with the compounds of theinvention.

Tumours with mutants of CDCl₄ or up-regulation, in particularover-expression, of cyclin E or loss of p21 or p27 may be particularlysensitive to CDK inhibitors. Tumours may preferentially be screened forup-regulation, in particular over-expression, of cyclin E (Harwell R M,Mull B B, Porter D C, Keyomarsi K.; J Biol Chem. 2004 Mar. 26;279(13):12695-705) or loss of p21 or p27 or for CDCl₄ variants prior totreatment (Rajagopalan H, Jallepalli P V, Rago C, Velculescu V E,Kinzler K W, Vogelstein B, Lengauer C.; Nature. 2004 Mar. 4;428(6978):77-81).

Patients with mantle cell lymphoma (MCL) could be selected for treatmentwith a compound of the invention using diagnostic tests outlined herein.MCL is a distinct clinicopathologic entity of non-Hodgkin's lymphoma,characterized by proliferation of small to medium-sized lymphocytes withco-expression of CD5 and CD20, an aggressive and incurable clinicalcourse, and frequent t(11;14)(q13;q32) translocation. Over-expression ofcyclin D1 mRNA, found in mantle cell lymphoma (MCL), is a criticaldiagnostic marker. Yatabe et al (Blood. 2000 Apr. 1; 95(7):2253-61)proposed that cyclin D1-positivity should be included as one of thestandard criteria for MCL, and that innovative therapies for thisincurable disease should be explored on the basis of the new criteria.Jones et al (J Mol Diagn. 2004 May; 6(2):84-9) developed a real-time,quantitative, reverse transcription PCR assay for cyclin D1 (CCND1)expression to aid in the diagnosis of mantle cell lymphoma (MCL). Howeet al (Clin Chem. 2004 January; 50(1):80-7) used real-time quantitativeRT-PCR to evaluate cyclin D1 mRNA expression and found that quantitativeRT-PCR for cyclin D1 mRNA normalized to CD19 mRNA can be used in thediagnosis of MCL in blood, marrow, and tissue. Alternatively, patientswith breast cancer could be selected for treatment with a CDK inhibitorusing diagnostic tests outline above. Tumour cells commonly overexpresscyclin E and it has been shown that cyclin E is over-expressed in breastcancer (Harwell et al, Cancer Res, 2000, 60, 481-489). Therefore breastcancer may in particular be treated with a CDK inhibitor as providedherein.

In addition, the cancer may be analysed for INK4a and RB loss offunction, and cyclin D1 or CDK4 overexpression or CDK4 mutation. RB lossand mutations inactivating p16^(INK4a) function or hypermethylation ofp16^(INK4a) occur in many tumour types. Rb is inactivated in 100%retinoblastomas and in 90% of small cell lung carcinomas. Cyclin D1 isamplified in 40% of head and neck, over-expressed in 50% of breastcancers and 90% of mantle cell lymphomas. p16 is deleted in 60% ofnon-small lung carcinomas and in 40% of pancreatic cancers. CDK4 isamplified in 20% of sarcomas and in 10% of gliomas. Events resulting inRB or p16^(INK4a) inactivation through mutation, deletion, or epigeneticsilencing, or in the overexpression of cyclin D1 or Cdk4 can beidentified by the techniques outlined herein. Tumours withup-regulation, in particular over-expression of cyclin D or CDK4 or lossof INK4a or RB may be particularly sensitive to CDK inhibitors. Thus,the patient may be subjected to a diagnostic test to detect a markercharacteristic of over-expression of cyclin D or CDK4 or loss of INK4aor RB.

Cancers that experience INK4a and RB loss of function and cyclin D1 orCDK4 overexpression, include small cell lung cancer, non-small cell lungcancer, pancreatic cancer, breast cancer, glioblastoma multiforme, Tcell ALL and mantle cell lymphoma. Therefore patients with small celllung cancer, non-small cell lung cancer, pancreatic cancer, breastcancer, glioblastoma multiforme, T cell ALL or mantle cell lymphomacould be selected for treatment with a CDK inhibitor using diagnostictests outlined above and may in particular be treated with a CDKinhibitor as provided herein.

Patients with specific cancers caused by aberrations in theD-Cyclin-CDK4/6-INK4-Rb pathway could be identified by using thetechniques described herein and then treated with a CDK4 inhibitor asprovided. Examples of abnormalities that activate or sensitise tumoursto CDK4 signal include, receptor activation e.g. Her-2/Neu in breastcancer, ras mutations for example in pancreatic, colorectal or lungcancer, raf mutations for example in melanoma, p16 mutations for examplein melanoma, p16 deletions for example in lung cancer, p16 methylationfor example in lung cancer or cyclin D overexpression for example inbreast cancer. Thus, a patient could be selected for treatment with acompound of the invention using diagnostic tests as outlined herein toidentify up-regulation of the D-Cyclin-CDK4/6-INK4-Rb pathway forexample by overexpression of cyclin D, mutation of CDK4, mutation ordepletion of pRb, deletion of p16-INK4, mutation, deletion ormethylation of p16, or by activating events upstream of the CDK4/6kinase e.g. Ras mutations or Raf mutations or hyperactive orover-expressed receptors such as Her-2/Neu.

The compounds of the present invention are particularly advantageous inthat they are selective inhibitors of CDK4 over other cyclin dependentkinases. PCT/US2007/069595 generically discloses compounds of thisclass, but the presently claimed compounds have increased potency andselectivity of CDK4 over other cyclin dependent kinases. This isadvantageous in developing a drug suitable for use as a CDK4 inhibitor.

More particularly and with regard to the generic application, thefollowing compounds (from PCT/US2007/069595) of table 3 represent theclosest prior art to the chemotype of the presently claimed invention:

TABLE 3 (Prior Art) Compound Example Number

200

201

202

The following table 4 shows the inhibition against relevant targets ofcompounds of the prior art as compared to compounds of the presentinvention:

TABLE 4 Compound Number IC50 (μM) Selectivity 200 (prior art) CDK4:0.005 CDK1: >1.6 CDK2: >1.4 201(prior art) CDK4: 0.11 CDK1: 7.5 CDK2:10.3 202 (prior art) CDK4: 2.5 CDK1: >15 CDK2: >15 74 of presentapplication CDK4: 0.01 Greater than 11,000 fold CDK1: 113 selectiveagainst CDK4 CDK2: 76 63 of present application CDK4: 0.008 CDK1: >15CDK2: >15 26 of present application CDK4: 0.026 CDK1: >15 CDK2: >15

The superior selectivity of the presently claimed compounds againstother CDK family members and other kinases means that, compared to othercompounds with less selectivity, the presently claimed compounds wouldhave reduced off target activities, and therefore less unpredictedtoxicity in cells. When looking at the results of cell cycle analysisperformed with the presently claimed compounds and compound 200 of theprior art, for example, it is clear that, while the presently claimedcompounds maintain exclusive G1 arrest even at 10 uM concentrations,compound 200 starts to induce G2/M phase blocks at 1 and 10 uMconcentrations, reflecting its off-target activities at higher than 1 uMconcentrations. Moreover, the inhibitory effects of the CDK4 inhibitorare absolutely dependent upon the presence of the Retinoblastoma protein(pRb). Activities in pRb negative cells for candidate CDK4 inhibitorsindicate that the compounds have off target activities and not asselective. Compared to the presently claimed series, which are inert inpRb negative cells, compound 200 does inhibit the cell proliferation ofpRb negative cells at high concentrations, illustrating its off targetactivities.

Still further, it has been shown that, while the activity of CDK4 is notrequired for normal fibroblast cell proliferation, the inhibition ofCDK1 is thought to be an undesirable effect. When dosed to animals,compared to the prior art, the presently claimed compounds are expectedto induce less cytotoxicity. Therefore, the presently claimed compoundsare a superior CDK4 inhibitor compared to those of the same scaffoldswith the same K4 potency but less selectivity against otherCDKs/kinases, as the compound should have a higher therapeutic indexthan the less selective ones.

In an other embodiment, the present invention comprises the followingcompounds:

-   7-Cyclopentyl-2-[5-(3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3d]pyrimidine-6-carbonitrile;-   7-Cyclopentyl-2-{5-[4-(2-fluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(4-dimethylamino-3,-   4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   2-[5-(4-Carbamoylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   2-{5[4-(2-Amino-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   2-[5-(3-Amino-pyrrolidin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-methoxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[4-(2-hydroxyethyl)-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-((R)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-((S)-3-methylpiperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(3-methylpiperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(3-hydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(pyrrolidine-1-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-((S)-2,3-dihydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(5-{4-[2-(2-hydroxyethoxy)-ethyl]-piperazin-1-yl}-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-hydroxy-1-methylethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{6-[4-(2-hydroxyethyl)-piperazin-1-yl]-pyridazin-3-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2,3-dihydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-((R)-2,3-dihydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(4-dimethylamino-3,4,5,6-tetrahydro-2H-[1,3]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile;-   7-Cyclopentyl-2-(3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(4-dimethylaminopiperidine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-((S)-3-methylpiperazin-1-ylmethyl)-pyridin-2-ylamino]-7Hpyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-((S)-2-hydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-((R)-2-hydroxypropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid methylamide;-   7-Cyclopentyl-2-[5-(4-isopropyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(4-isopropyl-piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(4-methyl-pentyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[6-(4-isopropyl-piperazin-1-yl)-pyridazin-3-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-hydroxy-2methylpropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(3,3-dimethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(3,8-diaza-bicyclo[3.2.1]oct-3-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(4-ethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(4-cyclopentyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-(1′-isopropyl-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[(R)-4-(2-hydroxyethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[(S)-4-(2-hydroxyethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-hydroxyethyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-dimethylaminoacetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(2-ethyl-butyl)piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   2-{5-[4-(2-Cyclohexyl-acetyl)piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-{5-[4-(3-cyclopentyl-propionyl)-piperazin-1-yl]-pyridin-2-ylamino}7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[5-(4-isobutylpiperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic    acid dimethylamide;-   {4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)pyridin-3-yl]-piperazin-1-yl}-acetic    acid methyl ester;-   7-Cyclopentyl-2-{5-[4-(2-isopropoxyethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7Hpyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   {4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)pyridin-3-yl]-piperazin-1-yl}-acetic    acid ethyl ester;-   4-(6-{7-Cyclopentyl-6-[(2-hydroxy-ethyl)methyl-carbamoyl]-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino}-pyridin-3-yl)piperazine-1-carboxylic    acid tert-butyl ester;-   7-Cyclopentyl-2-{5-[4-(2-methyl-butyl)piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   7-Cyclopentyl-2-[1′-(2-hydroxy-ethyl)-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic    acid dimethylamide;-   {4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]piperazin-1-yl}-acetic    acid; and-   2-{4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionic    acid; or pharmaceutically acceptable salts thereof.

Assays

The inhibition of protein kinase activity by the compounds of theinvention may be measured using a number of assays available in the art.Examples of such assays are described in the Exemplification sectionbelow.

Pharmaceutical Compositions

The language “effective amount” of the compound is that amount necessaryor sufficient to treat or prevent a protein kinase-associated disorder,e.g. prevent the various morphological and somatic symptoms of a proteinkinase-associated disorder, and/or a disease or condition describedherein. In an example, an effective amount of the compound of theinvention is the amount sufficient to treat a protein kinase-associateddisorder in a subject. The effective amount can vary depending on suchfactors as the size and weight of the subject, the type of illness, orthe particular compound of the invention. For example, the choice of thecompound of the invention can affect what constitutes an “effectiveamount.” One of ordinary skill in the art would be able to study thefactors contained herein and make the determination regarding theeffective amount of the compounds of the invention without undueexperimentation.

The regimen of administration can affect what constitutes an effectiveamount. The compound of the invention can be administered to the subjecteither prior to or after the onset of a protein kinase-associateddisorder. Further, several divided dosages, as well as staggereddosages, can be administered daily or sequentially, or the dose can becontinuously infused, or can be a bolus injection. Further, the dosagesof the compound(s) of the invention can be proportionally increased ordecreased as indicated by the exigencies of the therapeutic orprophylactic situation.

Compounds of the invention may be used in the treatment of states,disorders or diseases as described herein, or for the manufacture ofpharmaceutical compositions for use in the treatment of these diseases.Methods of use of compounds of the present invention in the treatment ofthese diseases, or pharmaceutical preparations having compounds of thepresent invention for the treatment of these diseases.

The language “pharmaceutical composition” includes preparations suitablefor administration to mammals, e.g., humans. When the compounds of thepresent invention are administered as pharmaceuticals to mammals, e.g.,humans, they can be given per se or as a pharmaceutical compositioncontaining, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) ofactive ingredient in combination with a pharmaceutically acceptablecarrier.

The phrase “pharmaceutically acceptable carrier” is art recognized andincludes a pharmaceutically acceptable material, composition or vehicle,suitable for administering compounds of the present invention tomammals. The carriers include liquid or solid filler, diluent,excipient, solvent or encapsulating material, involved in carrying ortransporting the subject agent from one organ, or portion of the body,to another organ, or portion of the body. Each carrier must be“acceptable” in the sense of being compatible with the other ingredientsof the formulation and not injurious to the patient. Some examples ofmaterials which can serve as pharmaceutically acceptable carriersinclude: sugars, such as lactose, glucose and sucrose; starches, such ascorn starch and potato starch; cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients, such as cocoabutter and suppository waxes; oils, such as peanut oil, cottonseed oil,safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols,such as propylene glycol; polyols, such as glycerin, sorbitol, mannitoland polyethylene glycol; esters, such as ethyl oleate and ethyl laurate;agar; buffering agents, such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol; phosphate buffer solutions; and other non-toxiccompatible substances employed in pharmaceutical formulations.

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like;oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, α-tocopherol, and the like; and metal chelating agents, such ascitric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaricacid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical, buccal, sublingual, rectal, vaginal and/or parenteraladministration. The formulations may conveniently be presented in unitdosage form and may be prepared by any methods well known in the art ofpharmacy. The amount of active ingredient that can be combined with acarrier material to produce a single dosage form will generally be thatamount of the compound that produces a therapeutic effect. Generally,out of one hundred percent, this amount will range from about 1 percentto about ninety-nine percent of active ingredient, preferably from about5 percent to about 70 percent, most preferably from about 10 percent toabout 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules and the like), theactive ingredient is mixed with one or more pharmaceutically acceptablecarriers, such as sodium citrate or dicalcium phosphate, and/or any ofthe following: fillers or extenders, such as starches, lactose, sucrose,glucose, mannitol, and/or silicic acid; binders, such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; humectants, such as glycerol; disintegratingagents, such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate; solutionretarding agents, such as paraffin; absorption accelerators, such asquaternary ammonium compounds; wetting agents, such as, for example,cetyl alcohol and glycerol monostearate; absorbents, such as kaolin andbentonite clay; lubricants, such a talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, andmixtures thereof; and coloring agents. In the case of capsules, tabletsand pills, the pharmaceutical compositions may also comprise bufferingagents. Solid compositions of a similar type may also be employed asfillers in soft and hard-filled gelatin capsules using such excipientsas lactose or milk sugars, as well as high molecular weight polyethyleneglycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions that can be used include polymeric substances andwaxes. The active ingredient can also be in micro-encapsulated form, ifappropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluent commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants that may berequired.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the activecompound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents that delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissue.

The preparations of the present invention may be given orally,parenterally, topically, or rectally. They are of course given by formssuitable for each administration route. For example, they areadministered in tablets or capsule form, by injection, inhalation, eyelotion, ointment, suppository, etc., administration by injection,infusion or inhalation; topical by lotion or ointment; and rectal bysuppositories. Oral and/or IV administration is preferred.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically acceptable dosage forms by conventional methodsknown to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient which is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compound employed, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound that is the lowest dose effective to producea therapeutic effect. Such an effective dose will generally depend uponthe factors described above. Generally, intravenous and subcutaneousdoses of the compounds of this invention for a patient, when used forthe indicated analgesic effects, will range from about 0.0001 to about100 mg per kilogram of body weight per day, more preferably from about0.01 to about 50 mg per kg per day, and still more preferably from about1.0 to about 100 mg per kg per day. An effective amount is that amounttreats a protein kinase-associated disorder.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms.

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical composition.

Synthetic Procedure

Compounds of the present invention are prepared from commonly availablecompounds using procedures known to those skilled in the art, includingany one or more of the following conditions without limitation:

Within the scope of this text, only a readily removable group that isnot a constituent of the particular desired end product of the compoundsof the present invention is designated a “protecting group,” unless thecontext indicates otherwise. The protection of functional groups by suchprotecting groups, the protecting groups themselves, and their cleavagereactions are described for example in standard reference works, such ase.g., Science of Synthesis: Houben-Weyl Methods of MolecularTransformation. Georg Thieme Verlag, Stuttgart, Germany. 2005. 41627 pp.(URL: http://www.science-of-synthesis.com (Electronic Version, 48Volumes)); J. F. W. McOmie, “Protective Groups in Organic Chemistry”,Plenum Press, London and New York 1973, in T. W. Greene and P. G. M.Wuts, “Protective Groups in Organic Synthesis”, Third edition, Wiley,New York 1999, in “The Peptides”; Volume 3 (editors: E. Gross and J.Meienhofer), Academic Press, London and New York 1981, in “Methoden derorganischen Chemie” (Methods of Organic Chemistry), Houben Weyl, 4thedition, Volume 15/I, Georg Thieme Verlag, Stuttgart 1974, in H.-D.Jakubke and H. Jeschkeit, “Aminosäuren, Peptide, Proteine” (Amino acids,Peptides, Proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel1982, and in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharideand Derivate” (Chemistry of Carbohydrates: Monosaccharides andDerivatives), Georg Thieme Verlag, Stuttgart 1974. A characteristic ofprotecting groups is that they can be removed readily (i.e., without theoccurrence of undesired secondary reactions) for example by solvolysis,reduction, photolysis or alternatively under physiological conditions(e.g., by enzymatic cleavage).Salts of compounds of the present invention having at least onesalt-forming group may be prepared in a manner known per se. Forexample, salts of compounds of the present invention having acid groupsmay be formed, for example, by treating the compounds with metalcompounds, such as alkali metal salts of suitable organic carboxylicacids, e.g., the sodium salt of 2-ethylhexanoic acid, with organicalkali metal or alkaline earth metal compounds, such as thecorresponding hydroxides, carbonates or hydrogen carbonates, such assodium or potassium hydroxide, carbonate or hydrogen carbonate, withcorresponding calcium compounds or with ammonia or a suitable organicamine, stoichiometric amounts or only a small excess of the salt-formingagent preferably being used. Acid addition salts of compounds of thepresent invention are obtained in customary manner, e.g., by treatingthe compounds with an acid or a suitable anion exchange reagent.Internal salts of compounds of the present invention containing acid andbasic salt-forming groups, e.g., a free carboxy group and a free aminogroup, may be formed, e.g., by the neutralisation of salts, such as acidaddition salts, to the isoelectric point, e.g., with weak bases, or bytreatment with ion exchangers.Salts can be converted in customary manner into the free compounds;metal and ammonium salts can be converted, for example, by treatmentwith suitable acids, and acid addition salts, for example, by treatmentwith a suitable basic agent.Mixtures of isomers obtainable according to the invention can beseparated in a manner known per se into the individual isomers;diastereoisomers can be separated, for example, by partitioning betweenpolyphasic solvent mixtures, recrystallisation and/or chromatographicseparation, for example over silica gel or by, e.g., medium pressureliquid chromatography over a reversed phase column, and racemates can beseparated, for example, by the formation of salts with optically puresalt-forming reagents and separation of the mixture of diastereoisomersso obtainable, for example by means of fractional crystallisation, or bychromatography over optically active column materials. Intermediates andfinal products can be worked up and/or purified according to standardmethods, e.g., using chromatographic methods, distribution methods,(re-) crystallization, and the like.

General Process Conditions

The following applies in general to all processes mentioned throughoutthis disclosure.The process steps to synthesize the compounds of the invention can becarried out under reaction conditions that are known per se, includingthose mentioned specifically, in the absence or, customarily, in thepresence of solvents or diluents, including, for example, solvents ordiluents that are inert towards the reagents used and dissolve them, inthe absence or presence of catalysts, condensation or neutralizingagents, for example ion exchangers, such as cation exchangers, e.g., inthe H⁺ form, depending on the nature of the reaction and/or of thereactants at reduced, normal or elevated temperature, for example in atemperature range of from about −100° C. to about 190° C., including,for example, from approximately −80° C. to approximately 150° C., forexample at from −80 to −60° C., at room temperature, at from −20 to 40°C. or at reflux temperature, under atmospheric pressure or in a closedvessel, where appropriate under pressure, and/or in an inert atmosphere,for example under an argon or nitrogen atmosphere.At all stages of the reactions, mixtures of isomers that are formed canbe separated into the individual isomers, for example diastereoisomersor enantiomers, or into any desired mixtures of isomers, for exampleracemates or mixtures of diastereoisomers, for example analogously tothe methods described in Science of Synthesis: Houben-Weyl Methods ofMolecular Transformation. Georg Thieme Verlag, Stuttgart, Germany. 2005.The solvents from which those solvents that are suitable for anyparticular reaction may be selected include those mentioned specificallyor, for example, water, esters, such as lower alkyl-lower alkanoates,for example ethyl acetate, ethers, such as aliphatic ethers, for examplediethyl ether, or cyclic ethers, for example tetrahydrofuran or dioxane,liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, suchas methanol, ethanol or 1- or 2-propanol, nitriles, such asacetonitrile, halogenated hydrocarbons, such as methylene chloride orchloroform, acid amides, such as dimethylformamide or dimethylacetamide, bases, such as heterocyclic nitrogen bases, for examplepyridine or N-methylpyrrolidin-2-one, carboxylic acid anhydrides, suchas lower alkanoic acid anhydrides, for example acetic anhydride, cyclic,linear or branched hydrocarbons, such as cyclohexane, hexane orisopentane, or mixtures of those solvents, for example aqueoussolutions, unless otherwise indicated in the description of theprocesses. Such solvent mixtures may also be used in working up, forexample by chromatography or partitioning.The compounds, including their salts, may also be obtained in the formof hydrates, or their crystals may, for example, include the solventused for crystallization. Different crystalline forms may be present.The invention relates also to those forms of the process in which acompound obtainable as an intermediate at any stage of the process isused as starting material and the remaining process steps are carriedout, or in which a starting material is formed under the reactionconditions or is used in the form of a derivative, for example in aprotected form or in the form of a salt, or a compound obtainable by theprocess according to the invention is produced under the processconditions and processed further in situ.

Prodrugs

This invention also encompasses pharmaceutical compositions containing,and methods of treating protein kinase-associated disorders throughadministering, pharmaceutically acceptable prodrugs of compounds of thecompounds of the invention. For example, compounds of the inventionhaving free amino, amido, hydroxy or carboxylic groups can be convertedinto prodrugs. Prodrugs include compounds wherein an amino acid residue,or a polypeptide chain of two or more (e.g., two, three or four) aminoacid residues is covalently joined through an amide or ester bond to afree amino, hydroxy or carboxylic acid group of compounds of theinvention. The amino acid residues include but are not limited to the 20naturally occurring amino acids commonly designated by three lettersymbols and also includes 4-hydroxyproline, hydroxylysine, demosine,isodemosine, 3-methylhistidine, norvalin, beta-alanine,gamma-aminobutyric acid, citrulline homocysteine, homoserine, ornithineand methionine sulfone. Additional types of prodrugs are alsoencompassed. For instance, free carboxyl groups can be derivatized asamides or alkyl esters. Free hydroxy groups may be derivatized usinggroups including but not limited to hemisuccinates, phosphate esters,dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs ofhydroxy and amino groups are also included, as are carbonate prodrugs,sulfonate esters and sulfate esters of hydroxy groups. Derivatization ofhydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers wherein theacyl group may be an alkyl ester, optionally substituted with groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities, or where the acyl group is an amino acid ester asdescribed above, are also encompassed. Prodrugs of this type aredescribed in J. Med. Chem. 1996, 39, 10. Free amines can also bederivatized as amides, sulfonamides or phosphonamides. All of theseprodrug moieties may incorporate groups including but not limited toether, amine and carboxylic acid functionalities.Any reference to a compound of the present invention is therefore to beunderstood as referring also to the corresponding pro-drugs of thecompound of the present invention, as appropriate and expedient.

Combinations

A compound of the present invention may also be used in combination withother agents, e.g., an additional protein kinase inhibitor that is or isnot a compound of the invention, for treatment of a proteinkinase-associated disorder in a subject.By the term “combination” is meant either a fixed combination in onedosage unit form, or a kit of parts for the combined administrationwhere a compound of the present invention and a combination partner maybe administered independently at the same time or separately within timeintervals that especially allow that the combination partners show acooperative, e.g., synergistic, effect, or any combination thereof.The compounds of the invention may be administered, simultaneously orsequentially, with an antiinflammatory, antiproliferative,chemotherapeutic agent, immunosuppressant, anti-cancer, cytotoxic agentor kinase inhibitor other than a compound of the Formula I or saltthereof. Further examples of agents that may be administered incombination with the compounds of the invention include, but are notlimited to, a PTK inhibitor, cyclosporin A, CTLA4-Ig, antibodiesselected from anti-ICAM-3, anti-IL-2 receptor, anti-CD45RB, anti-CD2,anti-CD3, anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody OKT3,agents blocking the interaction between CD40 and gp39, fusion proteinsconstructed from CD40 and gp39, inhibitors of NF-kappa B function,non-steroidal antiinflammatory drugs, steroids, gold compounds,antiproliferative agents, FK506, mycophenolate mofetil, cytotoxic drugs,TNF-α inhibitors, anti-TNF antibodies or soluble TNF receptor,rapamycin, leflunimide, cyclooxygenase-2 inhibitors, paclitaxel,cisplatin, carboplatin, doxorubicin, carminomycin, daunorubicin,aminopterin, methotrexate, methopterin, mitomycin C, ecteinascidin 743,porfiromycin, 5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosinearabinoside, podophyllotoxin, etoposide, etoposide phosphate,teniposide, melphalan, vinblastine, vincristine, leurosidine,epothilone, vindesine, leurosine, or derivatives thereof.The compound of the invention and any additional agent may be formulatedin separate dosage forms. Alternatively, to decrease the number ofdosage forms administered to a patient, the compound of the inventionand any additional agent may be formulated together in any combination.For example, the compound of the invention inhibitor may be formulatedin one dosage form and the additional agent may be formulated togetherin another dosage form. Any separate dosage forms may be administered atthe same time or different times.Alternatively, a composition of this invention comprises an additionalagent as described herein. Each component may be present in individualcompositions, combination compositions, or in a single composition.

EXEMPLIFICATION OF THE INVENTION

The invention is further illustrated by the following examples, whichshould not be construed as further limiting. The practice of the presentinvention will employ, unless otherwise indicated, conventionaltechniques of cell biology, cell culture, molecular biology, transgenicbiology, microbiology and immunology, which are within the skill of theart.

Experimental Procedure Analytical Methods

In the examples, the compounds prepared are characterized by liquidchromatography and mass spectroscopy using the systems and operatingconditions set out below. Where atoms with different isotopes arepresent and a single mass quoted, the mass quoted for the compound isthe monoisotopic mass (i.e. ³⁵Cl; ⁷⁹Br etc.). Several systems are used,as described below, and these are equipped with, and are set up to rununder, closely similar operating conditions. The operating conditionsused are also described below.

LCMS analysis is performed using the following methods:

Waters Platform LC-MS System: HPLC System: Waters 2795 Mass SpecDetector: Micromass Platform LC PDA Detector: Waters 2996 PDA

Purity is measured by UV diode array detector (210-340 nm)

Method A

Eluent A: H₂O (10 mM NH₄HCO₃ buffer adjusted to pH=9.2 with NH₄OH)

Eluent B: CH₃CN

Gradient: 05-95% eluent B over 15 minutesFlow: 0.8 ml/min

Column: Waters XBridge C18 5μ, 2.1×50 mm Method B

Eluent A: H₂O (10 mM NH₄HCO₃ buffer adjusted to pH=9.2 with NH₄OH)

Eluent B: CH₃CN

Gradient: 05-95% eluent B over 3.5 minutesFlow: 0.8 ml/min

Column: Waters XBridge C18 5μ, 2.1×50 mm Method C Eluent A: H₂O (0.1%Formic Acid) Eluent B: CH₃CN (0.1% Formic Acid)

Gradient: 5-95% eluent B over 3.5 minutesFlow: 0.8 ml/min

Column: Phenomenex Synergi 4μ MAX-RP 80A, 2.0×50 mm Method D Eluent A:H2O (0.1% Formic Acid) Eluent B: CH3CN (0.1% Formic Acid)

Gradient: 5-95% acetonitrile/water over 7.75 minutesFlow: 1.0 ml/minColumn: Inertsil ODS3 100×3 mm C18 column

Waters Fractionlynx LC-MS System:

HPLC System: 2767 autosampler-2525 binary gradient pump

Mass Spec Detector: Waters ZQ PDA Detector: Waters 2996 PDA

Purity is measured by UV diode array detector (200-340 nm)

Method E

Eluent A: H₂O (10 mM NH₄HCO₃ buffer adjusted to pH=9.2 with NH₄OH)

Eluent B: CH₃CN

Gradient: 05-95% eluent B over 3.5 minutesFlow: 2.0 ml/min

Method for Preparative Mass Directed Liquid Chromatography (LCMS) WatersFractionlynx System: 2767 Dual Loop Autosampler/Fraction Collector

2525 preparative pumpCFO (column fluidic organiser) for column selectionRMA (Waters reagent manager) as make up pump

Waters ZQ Mass Spectrometer

Waters 2996 Photo Diode Array detector

Waters ZQ Mass Spectrometer Software Masslynx 4.1 Waters MS RunningConditions

Capillary voltage: 3.5 kV (3.2 kV on ES Negative)Cone voltage: 25 V

Source Temperature: 120° C. Multiplier: 500 V Scan Range: 125-800 amuIonisation Mode: ElectroSpray Positive or

ElectroSpray Negative

Once the analytical trace showed good chromatography a suitablepreparative method of the same type is chosen. Typical running conditionis:

Column Waters XBridge C18 5μ 100×19 mm or Phenomenex Gemini, 5□,100×21.2 mm) Mobile Phase

Solvent A: H₂O+10 mM NH₄HCO₃+NH₄OH, pH=9.2

Solvent B: CH₃CN

Flow rate: 24 ml/minGradient: Generally all gradients have an initial 0.4 min step with 95%A+5% B. Then according to analytical trace a 3.6 min gradient is chosenin order to achieve good separation (e.g. from 5% to 50% B for earlyretaining compounds; from 35% to 80% B for middle retaining compoundsand so on).Ish: 1.2 minute ish step is performed at the end of the gradientRe-equilibration: 2.1 minutes re-equilibration step is ran to preparethe system for the next runMake Up flow rate: 1 ml/minAll compounds are usually dissolved in 100% MeOH or 100% DMSO

Experimental Procedures General Procedure A (BOC-Deprotection)

Starting material is treated with excess HCl (4M solution in dioxane).MeOH and/or CHCl₃ as added to aid dissolution where necessary. After 16h, the sample is evaporated in vacuo and the residue purified by eitherSiO₂ chromatography, ion exchange chromatography or preparative LCMS.

Nitrile Analogues Example A

To a stirred solution of 5-bromo-2-nitropyridine (4.93 g, 24.3 mmol) andpiperazine-1-carboxylic acid tert-butyl ester (4.97 g, 26.7 mmol) inCH₃CN (60 ml) is added DIPEA (4.65 mL, 26.7 mmol). The mixture is heatedat reflux for 72 hours then cooled to room temperature and theprecipitated product collected by filtration. The filtrate isconcentrated and purified by flash column chromatography eluting with30% EtOAc/petrol. The combined products are re-crystallized fromEtOAc/petrol to give 4-(6-nitro-pyridin-3-yl)-piperazine-1-carboxylicacid tert-butyl ester, (4.50 g, 80% yield). MS(ESI) m/z 308 (M+H)⁺

Example B

A mixture of 4-(6-nitro-pyridin-3-yl)-piperazine-1-carboxylic acidtert-butyl ester (3.40 g, 11.0 mmol) and 10% Pd—C (400 mg, 0.376 mmol)in ethanol (100 ml) and ethyl acetate (100 ml) is agitated under 1atmosphere pressure of hydrogen overnight. The mixture is filtered andconcentrated to give of 4-(6-amino-pyridin-3-yl)-piperazine-1-carboxylicacid tert-butyl ester (2.87 g, 94% yield). MS(ESI) m/z 278 (M+H)⁺

Example 1047-Cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile

To a stirred solution of (5-bromo-2-chloropyrimidin-4-yl)-cyclopentyl-amine (1.00 g, 3.62 mmol) andPdCl₂(dppf)⋅dichloromethane (148 mg, 0.181 mmol) in THF (10 mL) is addedEt₃N (0.757 mL, 5.43 mmol) and 3,3-diethoxy-propyne (0.778 mL, 5.43mmol) sequentially at room temperature. The mixture is degassed under astream of N₂ and stirred at room temperature for 10 minutes before CuI(29 mg, 0.154 mmol) is added. The reaction vessel is evacuated andback-filled with N₂ (×3) and heated at 60° C. for 48 hours. The mixtureis allowed to cool, diluted with EtOAc, filtered and partitioned betweenH₂O and ethyl acetate. The phases are separated and the aqueous layer isfurther extracted with EtOAc (×3), combined organic extracts are dried(MgSO₄), filtered and concentrated. The residue is purified by SiO₂chromatography, eluting with a gradient of 5% EtOAc/petrol to 20%EtOAc/petrol to give[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-cyclopentyl amine(636 mg, 54%). MS(ESI) m/z 324.2 (M+H)⁺

To a stirred solution of[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-cyclopentyl-amine(7.50 g, 23.3 mmol) in THF (45 mL) is added 1N TBAF in THF (100 mL, 116mmol) at room temperature. The reaction mixture is heated under refluxovernight. After cooling the mixture is partitioned between H₂O anddichloromethane. The phases are separated and the aqueous layer isextracted with dichloromethane (×2). The combined organic extracts aredried (MgSO₄), filtered and concentrated. The residue is purified bySiO₂ chromatography eluting with a gradient of 10% EtOAc/petrol to 30%EtOAc/petrol to give2-chloro-7-cyclopentyl-6-diethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine,(5.68 g, 76%). MS(ESI) m/z 324.1 (M+H)⁺

To a stirred solution of2-chloro-7-cyclopentyl-6-diethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine(6.29 g, 19.5 mmol) in 1,4-dioxane (68 mL) is added conc. HCl (19 mL) atroom temperature. The reaction mixture is stirred for 30 minutes, thenneutralized with 2N NaOH aqueous solution and saturated NaHCO₃ aqueoussolution. The mixture extracted into EtOAc (×3), combined organicextracts are dried (MgSO₄), filtered and concentrated to give 6 g of thecrude 2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehydeas a beige solid. To a stirred suspension of the crude2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde inMeCN (125 mL) and H₂O (125 mL) is added H₂N—SO₃H (6.62 g, 58.5 mmol) atroom temperature. The reaction mixture is stirred for 3 hours before thepH is made >10 with 2N NaOH aqueous solution and the reaction stirredfor 1 hour. The mixture extracted into dichloromethane (×3), combinedorganic extracts are dried (MgSO₄), filtered and concentrated. Theresidue is purified by SiO₂ chromatography, eluting with a gradient of5% EtOAc/petrol to 20% EtOAc/petrol providing 4.00 g of2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile as awhite solid, 83% yield. MS(ESI) m/z 247.0 (M+H)⁺

Buchwald Procedure A

To a stirred solution of2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile (80mg, 0.324 mmol) in toluene (5.0 mL) is added sequentially Pd₂(dba)₃ (16mg, 0.0162 mmol), 2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl(14 mg, 0.0324 mmol) and4-(6-amino-pyridin-3-yl)-piperazine-1-carboxylic acid tert-butyl ester(Example B) (99 mg, 0.357 mmol). The mixture is degassed under a streamof N₂ before LiHMDS (1M in THF; 0.650 mL, 0.650 mmol) is added. Thereaction mixture is heated at 110° C. overnight. At room temperature themixture is diluted with EtOAc filtered and concentrated. The residue ispurified by SiO₂ chromatography, eluting with EtOAc gave 35 mg ofmaterial which is triturated with a 1:1 mixture of EtOAc/petrolproviding4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazine-1-carboxylicacid tert-butyl ester (20 mg).

Using General Procedure A,4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazine-1-carboxylicacid tert-butyl ester (20 mg) gave crude product which is purified bySCX column (eluting with a 1:17 mixture of 2M NH₃ inMeOH/dichloromethane) to give a solid. Trituration with diethyl ethergave7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile(8.8 mg, 7%) (over 2 steps). MS(ESI) m/z 389.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, DMSO-d₆): 9.68 (1H, s), 8.91 (1H, s), 8.11 (1H, d),8.01 (1H, d), 7.51 (1H, s), 7.43 (1H, dd), 5.07 (1H, quintet), 3.10-2.99(4H, m), 2.92-2.78 (4H, m), 2.32-2.08 (4H, m), 2.08-1.92 (2H, m),1.82-1.66 (2H, m).

Example 477-Cyclopentyl-2-((4-dimethylaminopiperidin)-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile

By repeating procedures described in Example A,4-dimethylaminopiperidine (2.60 g, 18.4 mmol) to givedimethyl-[1-(6-nitro-pyridin-3-yl)-piperidin-4-yl]-amine, (3.90 g, 80%)(purification by precipitation). MS(ESI) m/z 250.1 (M+H)⁺

By repeating procedures described Example B,dimethyl-[1-(6-nitro-pyridin-3-yl)-piperidin-4-yl]-amine (3.90 g, 15.6mmol) gave 5-(4-dimethylaminopiperidin-1-yl)-pyridin-2-ylamine (3.32 g,97%). [M−H]⁺=219.1.

Following Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile (95mg, 0.385 mmol) and 5-(4-dimethylaminopiperidin-1-yl)-pyridin-2-ylamine(93 mg, 0.424 mmol) to give7-cyclopentyl-2-((4-dimethylaminopiperidin)-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile(77 mg, 46%) [following trituration with 1:1 mixture of EtOAc/petrol]

MS(ESI) m/z 431.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, DMSO-d₆): 9.66 (1H, s), 8.90 (1H, s), 8.09 (1H, d),8.02 (1H, d), 7.51 (1H, s), 7.45 (1H, dd), 5.07 (1H, quintet), 3.74-3.62(2H, m), 2.75-2.63 (2H, m), 2.30-2.08 (11H, m), 2.08-1.92 (2H, m),1.92-1.81 (2H, m), 1.81-1.66 (2H, m), 1.59-1.44 (2H, m).

Example 2rac-7-Cyclopentyl-2-[5-(3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile

By repeating procedures described in Example A,2-methyl-piperazine-1-carboxylic acid tert-butyl ester (1.08 g, 5.40mmol) gave 2-methyl-4-(6-nitro-pyridin-3-yl)-piperazine-1-carboxylicacid tert-butyl ester (0.610 g, 39%) (following SiO₂ chromatography,eluting with 2% MeOH/dichloromethane). MS(ESI) m/z 323 (M+H)⁺

By repeating procedures described in Example B,2-methyl-4-(6-nitro-pyridin-3-yl)-piperazine-1-carboxylic acidtert-butyl ester (600 mg, 1.52 mmol) is hydrogenated over Pd—C on anH-cube (Thales) (instead of under an atmosphere of hydrogen) to give4-(6-amino-pyridin-3-yl)-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (544 mg, 98%). MS(ESI) m/z 293 (M+H)⁺

Using Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile (95mg, 0.385 mmol) and4-(6-amino-pyridin-3-yl)-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (124 mg, 0.424 mmol) gave4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (128 mg) [following SiO₂ chromatography elutingwith 1-2.5% MeOH/dichloromethane and subsequent trituration with diethylether]. The material is used directly in the next step.

Using General Procedure A,4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester gave7-cyclopentyl-2-[5-(3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile(91 mg, 59% over 2 steps) [following purification by Strata-NH₂ column,eluting with a 1:1 mixture of MeOH/dichloromethane, and subsequenttrituration with diethyl ether]. MS(ESI) m/z 403.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, DMSO-d₆): 10.00 (1H, s), 9.00-8.87 (2H, m), 8.64-8.51(1H, m), 8.17-8.05 (2H, m), 7.61 (1H, d), 7.55 (1H, s), 5.10 (1H,quintet), 3.85-3.67 (2H, m), 3.50-3.36 (2H, m), 3.20 (1H, dd), 2.97 (1H,t), 2.75 (1H, t), 2.32 (3H, s), 2.29-2.10 (4H, m), 2.08-1.94 (2H, m),1.82-1.68 (2H, m), 1.29 (3H, d).

Example 1067-Cyclopentyl-2-(5-[1,4]diazepan-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile

By repeating procedures described in Example A,[1,4]diazepane-1-carboxylic acid tert-butyl ester (1.08 g, 5.40 mmol) inCH₃CN (20 ml) gave 4-(6-nitro-pyridin-3-yl)-[1,4]diazepane-1-carboxylicacid tert-butyl ester (533 mg) [following SiO₂ chromatography elutingwith 2% MeOH/dichloromethane]. MS(ESI) m/z 323 (M+H)⁺

By repeating procedures described in Example B,4-(6-nitro-pyridin-3-yl)-[1,4]diazepane-1-carboxylic acid tert-butylester (490 mg, 1.52 mmol) is hydrogenated over Pd—C on an H-cube(Thales) (instead of under an atmosphere of hydrogen) to give4-(6-amino-pyridin-3-yl)-[1,4]diazepane-1-carboxylic acid tert-butylester (544 mg, 98%). MS(ESI) m/z 293 (M+H)⁺

Following Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile (95mg, 0.385 mmol) and 4-(6-amino-pyridin-3-yl)-[1,4]diazepane-1-carboxylicacid tert-butyl ester (124 mg, 0.424 mmol) gave4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-[1,4]diazepane-1-carboxylicacid tert-butyl ester (96 mg) [following SiO₂ chromatography, elutingwith a 1-3% MeOH/dichloromethane and subsequent trituration with adiethyl ether]. The material is used directly in the next step.

Following General Procedure A,4-[6-(6-cyano-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-[1,4]diazepane-1-carboxylicacid tert-butyl ester gave7-cyclopentyl-2-(5-[1,4]diazepan-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile(71 mg, 46% over 2 steps [following purification by Strata-NH₂ column,eluting with a 1:1 mixture of MeOH/dichloromethane, and subsequenttrituration with diethyl ether]. MS(ESI) m/z 403.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, DMSO-d₆): 9.01 (1H, s), 8.67 (2H, s), 7.95-7.80 (2H,m), 7.73-7.54 (2H, m), 5.14 (1H, quintet), 3.75 (2H, t), 3.54 (2H, t),3.34-3.26 (2H, m), 3.23-3.14 (2H, m), 2.33 (3H, s), 2.27-2.13 (4H, m),2.13-1.93 (4H, m), 1.83-1.67 (2H, m).

Example 1057-Cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile

Following Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile (95mg, 0.385 mmol) and5-(tert-butyl-dimethyl-silanyloxymethyl)-pyridin-2-ylamine (101 mg,0.424 mmol) (Example C) gave 114 mg of2-[5-(tert-butyl-dimethyl-silanyloxymethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile[following SiO₂ chromatography eluting with 1-2% MeOH/dichloromethaneand subsequent trituration with diethyl ether].

To a stirred solution of2-[5-(tert-butyl-dimethyl-silanyloxymethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrilein THF (2.0 mL) is added HF⋅pyridine (0.080 mL) at 0° C. The reactionmixture stirred at room temperature for 16 h before it is diluted withethyl acetate, ished with saturated NaHCO₃ solution, dried (MgSO₄),filtered and concentrated. Trituration with diethyl ether gave7-cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carbonitrile,68 mg, 53% yield (over 2 steps). MS(ESI) m/z 335.0 (M+H)⁺ (method A).

¹H NMR (400 MHz, DMSO-d₆): 9.96 (1H, s), 8.97 (1H, s), 8.33-8.21 (2H,m), 7.75 (1H, dd), 7.55 (1H, s), 5.19 (1H, t), 5.10 (1H, quintet), 4.49(2H, d), 2.37-2.10 (4H, m), 2.10-1.92 (2H, m), 1.85-1.67 (2H, m).

Example 92-{5-[4-(2-Cyano-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example A, (except heating at 130°C. for 1 h in a CEM Discovery microwave, rather than heating at reflux)3-piperazin-1-yl-propionitrile (510 mg, 3.63 mmol) gave3-[4-(6-nitro-pyridin-3-yl)-piperazin-1-yl]-propionitrile as a whitecrystalline solid (212 mg, 25%) [following SiO₂ chromatography, elutingwith 0-10% methanol/dichlomethane and subsequent recrystallization formethyl acetate/petroleum ether (212 mg, 25%). MS(ESI) m/z 262.1 (M+H)⁺

By repeating procedures described in Example B,3-[4-(6-nitro-pyridin-3-yl)-piperazin-1-yl]-propionitrile (200 mg, 0.763mmol) gave 3-[4-(6-amino-pyridin-3-yl)-piperazin-1-yl]-propionitrile(165 mg, 94%) which is used in the next step without furtherpurification. MS(ESI) m/z 232.1 (M+H)⁺

Following Buchwald Procedure A,3-[4-(6-amino-pyridin-3-yl)-piperazin-1-yl]-propionitrile (173 mg, 0.751mmol) and2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (200 mg, 0.683 mmol) gave2-{5-[4-(2-cyano-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (34 mg, 10%) [following purification by preparativeLCMS]. MS(ESI) m/z 488.2 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d₃-OD): 8.72 (1H, s), 8.24 (1H, d), 7.99 (1H, d),7.51 (1H, dd), 6.62 (1H, s), 4.82-4.72 (1H, m), 3.23 (4H, t), 3.17 (6H,s), 2.82-2.65 (8H, m), 2.62-2.48 (2H, m), 2.17-1.98 (4H, m), 1.86-1.64(2H, m).

Example 257-Cyclopentyl-2-{5-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example A (except heating at 130°C. for 1 h in a CEM Discovery microwave, rather than heating at reflux),1-(2,2,2-trifluoroethyl)piperazine (1.31 g, 5.41 mmol) gave1-(6-nitro-pyridin-3-yl)-4-(2,2,2-trifluoro-ethyl)-piperazine (210 mg,15%) [following purification by SiO₂ chromatography, eluting with 0-10%MeOH/dichloromethane]. MS(ESI) m/z 291.1 (M+H)⁺

By repeating procedures described in Example B,6-nitro-pyridin-3-yl)-4-(2,2,2-trifluoro-ethyl)-piperazine (210 mg,0.724 mmol) gave5-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamine (158 mg,84%) which is used in the next step without further purification.MS(ESI) m/z 261.1 (M+H)⁺

Buchwald Method B

A mixture of5-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamine (158 mg,0.607 mmol),2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (118 mg, 0.405 mmol), Pd₂(dba)₃ (18.5 mg, 0.020 mmol),BINAP (25 mg, 0.040 mmol) and sodium-tert-butoxide (70 mg, 0.728 mmol)in dioxane (3.5 mL) is degassed and heated to 100° C. for 1 h in a CEMDiscover microwave. The reaction mixture is partitioned betweendichloromethane and saturated NaHCO₃ solution. The organic layer isseparated and the aqueous layer extracted with further dichloromethane.The combined organics are ished with brine, dried (MgSO₄), filtered andconcentrated. The crude product is purified using silica gelchromatography (0 to 10% methanol/dichloromethane) to give7-cyclopentyl-2-{5-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide, which is purified further by trituration withacetonitrile (115 mg, 55%). MS(ESI) m/z 517.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, Me-d₃-OD): 8.72 (1H, s), 8.24 (1H, d), 7.98 (1H, d),7.50 (1H, dd), 6.62 (1H, s), 4.81-4.72 (1H, m), 3.27-3.09 (12H, m), 2.89(4H, t), 2.61-2.49 (2H, m), 2.16-2.01 (4H, m), 1.81-1.69 (2H, m).

Example 87-Cyclopentyl-2-(5-[1,4]diazepan-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Using Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethyl amide (0.13 g, 0.444 mmol) gave4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-[1,4] diazepane-1-carboxylic acidtert-butyl ester (66 mg, 27%) [following purification by SiO₂chromatography eluting with 0-3% MeOH in dichloromethane]. MS(ESI) m/z549.3 (M+H)⁺

Using General Procedure A,4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-[1,4] diazepane-1-carboxylic acidtert-butyl ester (66 mg, 0.12 mmol) is used to give7-cyclopentyl-2-(5-[1,4]diazepan-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (35 mg, 65%) as a yellowish solid [followingpurification by SCX column chromatography eluting with 15% (NH₃ 2 M inMeOH)/DCM]. MS(ESI) m/z 449.2 (M+H)⁺ (method C).

¹H NMR (400 MHz, Me-d3-OD): 8.69 (1H, s), 8.08 (1H, d), 7.83 (1H, d),7.30 (1H, dd), 6.60 (1H, s), 4.76 (1H, quintet), 3.63 (4H, t), 3.17 (7H,s), 3.09 (2H, t), 2.91 (2H, t), 2.61-2.45 (2H, m), 2.17-1.93 (7H, m),1.80-1.63 (2H, m).

Example 13Rac-2-[5-(3-Amino-pyrrolidin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

By repeating procedures described in Example A, pyrrolidin-3-yl-carbamicacid tert-butyl ester (2.52 g, 13.5 mmol) gave[1-(6-nitro-pyridin-3-yl)-pyrrolidin-3-yl]-carbamic acid tert-butylester as a yellow solid (2.16 g, 57%) [following trituration with EtOAc]

[M+H]⁺=309.2.

By repeating procedures described in Example B,[1-(6-nitro-pyridin-3-yl)-pyrrolidin-3-yl]-carbamic acid tert-butylester (2.16 g, 7.01 mmol) gave[1-(6-amino-pyridin-3-yl)-pyrrolidin-3-yl]-carbamic acid tert-butylester as a purple solid (1.12 g, 56%). [following SiO₂ chromatographyeluting with 2.5-7.5% MeOH/dichloromethane). [M+H]⁺=279.2.

Using Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethyl amide (0.13 g, 0.444 mmol) and[1-(6-amino-pyridin-3-yl)-pyrrolidin-3-yl]-carbamic acid tert-butylester (0.136 g, 0.488 mmol) gave{1-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-pyrrolidin-3-yl}-carbamicacid tert-butyl ester (35 mg, 15%) (following SiO₂ chrlomatography,eluting with 0-3% MeOH/dichloromethane). MS(ESI) m/z 535.3 (M+H)⁺

Using General Procedure A,{1-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-pyrrolidin-3-yl}-carbamicacid tert-butyl ester (35 mg, 0.0655 mmol) gaverac-2-[5-(3-amino-pyrrolidin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide as a yellow solid (11 mg,39%) [following SiO₂ chromatography eluting with 5% (2.0 M NH₃ inMeOH)/DCM]. MS(ESI) m/z 435.2 (M+H)⁺ (method C).

¹H NMR (400 MHz, Me-d3-OD): 8.68 (1H, s), 8.10 (1H, d), 7.66 (1H, d),7.11 (1H, dd), 6.60 (1H, s), 4.79-4.66 (1H, m), 3.76-3.65 (1H, m),3.60-3.46 (2H, m), 3.17 (6H, s), 3.15-2.87 (2H, m), 2.62-2.44 (2H, m),2.37-2.22 (1H, m), 2.17-1.98 (4H, m), 1.98-1.80 (1H, m), 1.80-1.63 (2H,m).

Example 197-Cyclopentyl-2-[5-(3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Using Buchwald Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethyl amide (0.142 g, 0.485 mmol) and(+/−)-4-(6-amino-pyridin-3-yl)-2-methyl-piperazine-1-carboxylic acidtert-butyl ester (0.156 g, 0.533 mmol) gave4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (260 mg, 97%) (following SiO₂ chromatography,eluting with 0-3% MeOH/dichloromethane). MS(ESI) m/z 549.3 (M+H)⁺

Using General Procedure A,4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (260 mg, 0.474 mmol) gave7-cyclopentyl-2-[5-(3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxy-lic acid dimethylamide as a beige solid (67 mg,31%) [following SiO₂ chromatography eluting with 5% (2.0 M NH₃ inmethanol/dichloromethane]. MS(ESI) m/z 449.4 (M+H)⁺ (method D).

¹H NMR (400 MHz, DMSO-d6): 9.23 (1H, s), 8.76 (1H, s), 8.13 (1H, d),7.98 (1H, d), 7.41 (1H, dd), 6.60 (1H, s), 4.80-4.67 (1H, m), 3.46 (2H,t), 3.06 (6H, s), 3.02-2.90 (1H, m), 2.90-2.74 (2H, m), 2.61-2.49 (2H,m), 2.49-2.27 (2H, m), 2.27-2.08 (2H, m), 1.98 (4H, s), 1.65 (2H, d),1.03 (3H, d).

Example 57-Cyclopentyl-2-{5-[4-(2-fluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (150 mg, 0.318 mmol) and potassiumcarbonate (132 mg, 0.955 mmol) in acetonitrile (3 mL) and DMF (2 mL) isadded 1-bromo-2-fluoroethane (0.035 mL, 0.478 mmol) and the reactionmixture is heated to 80° C. for 24 h in a sealed reaction vial. Uponcooling, the reaction mixture is partitioned between dichloromethane andwater. The organic layer is separated and the aqueous layer extractedfurther with dichloromethane. The combined organics are ished withbrine, dried (MgSO₄), filtered and concentrated. The crude product ispurified using silica gel chromatography (0 to 10%methanol/dichloromethane) to give7-cyclopentyl-2-{5-[4-(2-fluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (84 mg, 55%) as an off white solid. MS(ESI) m/z 481.2(M+H)⁺ (method B).

¹H NMR (400 MHz, DMSO-d6): 9.23 (1H, s), 8.75 (1H, s), 8.14 (1H, d),7.99 (1H, d), 7.43 (1H, dd), 6.60 (1H, s), 4.79-4.68 (1H, m), 4.64 (1H,t), 4.53 (1H, t), 3.14 (4H, t), 3.06 (6H, s), 2.72 (1H, t), 2.68-2.60(5H, m), 2.49-2.37 (2H, m), 1.98 (4H, s), 1.65 (2H, d).

Example 842-[5-(4-Cyanomethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example 5,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (150 mg, 0.318 mmol) gave2-[5-(4-cyanomethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (99 mg, 66%). MS(ESI) m/z 474.4(M+H)⁺ (method B).

¹H NMR (400 MHz, DMSO-d₆): 9.25 (1H, s), 8.75 (1H, s), 8.15 (1H, d),8.04-7.97 (1H, d), 7.48-7.41 (1H, dd), 6.59 (1H, s), 4.78-4.69 (1H, m),3.81 (2H, s), 3.18 (4H, t), 3.06 (6H, s), 2.66 (5H, t), 2.43 (1H, d),1.99 (4H, s), 1.70-1.61 (2H, m).

Example 147-Cyclopentyl-2-{5-[4-(2-methoxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example 5,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (150 mg, 0.318 mmol) gave7-cyclopentyl-2-{5-[4-(2-methoxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (46 mg, 29%). MS(ESI) m/z 493.5(M+H)⁺ (method B).

¹H NMR (400 MHz, DMSO-d₆): 9.22 (1H, s), 8.75 (1H, s), 8.14 (1H, d),7.98 (1H, d), 7.42 (1H, dd), 6.60 (1H, s), 4.78-4.69 (1H, m), 3.48 (2H,t), 3.26 (3H, s), 3.18-3.09 (4H, m), 3.06 (6H, s), 2.65-2.52 (6H, m),2.47-2.35 (2H, m), 1.98 (4H, s), 1.71-1.58 (2H, m).

Example 102-[5-(4-Carbamoylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example 5,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (100 mg, 0.212 mmol) gave2-[5-(4-carbamoylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (66 mg, 63%) as an off white solid [following SiO₂chromatography eluting with 0 to 10% (2M NH₃ inmethanol/dichloromethane). MS(ESI) m/z 492.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, DMSO-d₆): 9.22 (1H, s), 8.75 (1H, s), 8.14 (1H, d),7.99 (1H, d), 7.43 (1H, dd), 7.25-7.17 (1H, m), 7.17-7.10 (1H, m), 6.59(1H, s), 4.77-4.70 (1H, m), 3.17 (4H, t), 3.06 (6H, s), 2.94 (2H, s),2.61 (4H, t), 2.44 (2H, s), 1.98 (4H, s), 1.65 (2H, d).

Example 337-Cyclopentyl-2-(5-{4-[2-(2-hydroxy-ethoxy)-ethyl]-piperazin-1-yl}-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a stirred solution of tert-butyldimethylchlorosilane (50% wt solutionin toluene, 8.38 mL, 24.08 mmol) and imidazole (1.78 g, 26.09 mmol) inDMF (10 mL) at 0° C. is added 2-(2-chloroethoxy)ethanol (2.13 mL, 20.07mmol) dropwise. The reaction mixture is then stirred for 1 h at 0° C.before warming to room temperature and stirring for a further 17 h. Thereaction mixture is partitioned between diethyl ether and brine. Thecombined organics are then dried (MgSO₄), filtered and concentratedunder reduced pressure to give product (76 mg, 0.318 mmol) which is useddirectly without further purification.

By repeating procedures described in Example 5,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (100 mg, 0.212 mmol) gave7-cyclopentyl-2-(5-{4-[2-(2-hydroxy-ethoxy)-ethyl]-piperazin-1-yl}-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (13 mg, 12%). MS(ESI) m/z 523.5(M+H)⁺ (method D).

¹H NMR (400 MHz, DMSO-d₆): 9.22 (1H, s), 8.75 (1H, s), 8.14 (1H, d),7.99 (1H, d), 7.43 (1H, dd), 6.60 (1H, s), 4.79-4.68 (1H, m), 4.61 (1H,t), 3.57 (2H, t), 3.50 (2H, q), 3.44 (2H, t), 3.15-3.09 (4H, m), 3.06(6H, s), 2.60 (4H, t), 2.54 (2H, t), 2.48-2.37 (2H, m), 2.05-1.91 (4H,m), 1.71-1.59 (2H, m).

Example 887-Cyclopentyl-2-{5-[4-(2-dimethylamino-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (80 mg, 0.170 mmol),N,N-dimethylglycine (18 mg, 0.170 mmol) and diisopropylethylamine (0.089mL, 0.509 mmol) in DMF (1 mL) is added TBTU (55 mg, 0.170 mmol) and thereaction mixture is stirred at room temperature for 1 hour. Methanol(0.5 mL) is added and the reaction mixture purified by silica gelchromatography (gradient of 0-10% 2M NH₃ in methanol/dichloromethane) togive7-cyclopentyl-2-{5-[4-(2-dimethylamino-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide, which is further purified by trituration withacetonitrile, as an off white solid (69 mg, 78%). MS(ESI) m/z 520.4(M+H)⁺ (method D).

¹H NMR (400 MHz, Me-d₃-OD): 8.75 (1H, s), 8.28-8.21 (1H, m), 8.01 (1H,d), 7.61-7.55 (1H, m), 6.65 (1H, s), 4.83-4.75 (1H, m), 4.22 (2H, s),3.84 (2H, t), 3.64 (2H, t), 3.28-3.21 (4H, m), 3.18 (6H, s), 2.92 (6H,s), 2.60-2.50 (2H, m), 2.16-2.02 (4H, m), 1.80-1.70 (2H, m).

Example 122-{5-[4-(2-Amino-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures described in Example 88,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide hydrochloride (150 mg, 0.318 mmol), N-BOC-glycine (56mg, 0.318 mmol) gave a crude product which is purified by SiO₂chromatography, eluting with 0-7% methanol/dichloromethane to give(2-{4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-2-oxo-ethyl)-carbamicacid tert-butyl ester which is used directly in the next step.

Following General Procedure A,(2-{4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-2-oxo-ethyl)-carbamicacid tert-butyl ester gave2-{5-[4-(2-amino-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as a pale yellow solid (96 mg, 61%) [followingpurification by SiO₂ chromatography eluting with 0-10% (2M NH₃ inmethanol/dichloromethane]. MS(ESI) m/z 492.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d₃-OD): 8.73 (1H, s), 8.28 (1H, d), 8.02 (1H, d),7.55 (1H, dd), 6.64 (1H, s), 4.82-4.73 (1H, m), 3.97 (2H, s), 3.88-3.78(2H, m), 3.65 (2H, t), 3.28-3.19 (4H, m), 3.17 (6H, s), 2.55 (2H, d),2.09 (4H, m), 1.82-1.69 (2H, m).

Benzylic Amine Analogues General Procedure B (Na(OAc)₃BH ReductiveAmination)

7-Cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (1 mol. eq.), amine (1.1 mol eq.) are dissolved indichloromethane (˜30 vols.) and stirred until the solution is clear (incases where the amine is sourced as a hydrochloride salt, 1 mol. eq.Et₃N is added). Where necessary, MeOH and/or 1 drop acetic acid is addedto aid dissolution and imine formation. Na(OAc)₃BH (1.5-2 mol. eq.) isthen added to the mixture and stirring continued at rt for further 16 h.The reaction is quenched with aqueous NaHCO₃ solution and the productextracted with dichloromethane, ethyl acetate or CHCl₃/i-PrOH (2:1). Thecombined organic fractions are dried (Na₂SO₄ or MgSO₄) filtered and thesolvent evaporated. The crude product is purified by SiO₂chromatography.

General Procedure C (NaCNBH₃ or NaBH₄ Reductive Amination)

7-Cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (1 mol. eq.) and amine (1-2 mol. eq.) are dissolvedin either dichloroethane/THF (3:1) or MeOH/dichloromethane mixtures (40vols.). The mixture is stirred at 20-40° C. for 16 hours, then cooled to0° C., NaCNBH₃ or NaBH₄ (1.5-2 mol. eq.) are then added and the mixturestirred at rt for 5 h. Where necessary, further MeOH and/or acetic acidis added to aid reaction progress. The mixture is then quenched withaqueous NaHCO₃ solution (10 ml) and the product extracted with eitherdiethyl ether, dichloromethane or CHCl₃/iPrOH (1:1). The combinedorganics are dried (MgSO₄), filtered and the solvent evaporated. Thecrude product is purified by SiO₂ chromatography.

Example C 5-(tert-Butyl-dimethyl-silanyloxymethyl)-pyridin-2-ylamine

To a solution of (6-chloro-3-pyridinyl) methanol (12.5 g, 87 mmol) andimidazole (7.2 g, 105 mmol) in THF (120 ml) is added a solution ofTBDMSCl (15.8 g, 105 mmol) in THF (60 ml). The mixture is stirred at rtfor 5 h and then is concentrated in vacuo to ¼ of the original volume.The slurry is then partitioned between water (60 ml) and EtOAc (60 ml).The organic layer is ished once with water, once with a 5% KH₂PO₄solution, once with sat. NaHCO₃ and finally once with brine. It is thendried (MgSO₄), filtered and the solvent evaporated in vacuo.5-(tert-butyl dimethyl silanyloxymethyl)-2-chloro pyridine is obtainedas a colorless liquid (22.4 g, 83%). MS(ESI) m/z 258.0 (M+H)⁺

5-(tert-butyl dimethyl silanyloxymethyl)-2-chloro pyridine (5.58 g, 21.6mmol), BINAP (0.4 g, 0.64 mmol) and benzophenone imine (4.7 g, 25.9mmol) are dissolved in toluene (50 ml) and the solution is degassed withnitrogen. Sodium t-butoxide (2.91 g, 30.3 mmol) and Pd₂(dba)₃ (0.2 g,0.22 mmol) are added and the solution is degassed once more. The mixtureis heated at 80° C. for 6 h and then allowed to cool to rt. The mixtureis diluted 4-fold with Et₂O and then the solution is filtered.Evaporation of solvent in vacuo gave the crudebenzhydrylidene[5-(tert-butyl dimethylsilanyloxymethyl)-pyridin-2-yl]-amine. This product is dissolved in MeOH(50 ml) and hydroxylamine (2.85 ml of a 50% aq. Solution, 46.5 mmol) andthe solution stirred at rt for 16 h. The mixture is concentrated invacuo and then the residue is dissolved in Et₂O (50 ml) and filtered.The filtrate is ished with brine, dried (MgSO₄) and then the solvent isevaporated in vacuo. The crude red oil is purified by SiO₂chromatography (gradient Petrol Ether:EtOAc=8:1 to 100% EtOAc) to givean orange oil which crystallized when triturated with hexane. Thesolvent is finally removed in vacuo to give 5-(tert-butyl dimethylsilanyloxymethyl)-pyridin-2-ylamine as an orange solid (2.9 g, 56.1%).MS(ESI) m/z 239.2 (M+H)⁺

Example 1087-Cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Following Buchwald Method B, 5-(tert-butyl dimethylsilanyloxymethyl)-pyridin-2-ylamine (0.980 g, 4.098 mmol) and2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethyl amide gave 2-[5-(tert-butyldimethyl-silanyloxymethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic acid dimethylamide (1.136 g, 84%) [followingSiO₂ chromatography eluting with 0-5% MeOH/dichloromethane). MS(ESI) m/z495.3 (M+H)⁺

2-[5-(tert-butyl dimethylsilanyloxymethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3d]pyrimidine-6-carboxylic acid dimethylamide (0.1 g, 0.202 mmol) isdissolved in dry THF (1 ml). TBAF (1M solution in THF) (0.303 ml, 0.303mmol) is added dropwise, then the mixture stirred for 16 h at rt. Thesolvent is then evaporated and the crude product is purified by SiO₂chromatography (eluting with 0-10% MeOH/dichloromethane) to give7-cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide (60 mg, 78%). MS(ESI) m/z381.2 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.77 (1H, s), 8.45 (1H, d), 8.25 (1H, d),7.82 (1H, dd), 6.65 (1H, s), 4.81-4.75 (1H, m), 4.62 (2H, s), 3.18 (6H,s), 2.65-2.53 (2H, m), 2.11 (4H, d), 1.84-1.72 (2H, m).

Example 1077-Cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

MnO₂ Method

7-Cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (4.49 g, 11.8 mmol) is dissolved in dichloromethane(175 ml) and methanol (75 ml). Activated MnO₂ 85% (51.1 g, 503 mmol) isadded in 4 portions over period of 48 h with continual stirring. After afurther 16 hours the mixture is filtered. The filtrate is heated to 38°C. and more MnO₂ (24 g, 236 mmol) is added as 2 batches over 5 hours.After stirring for a further 12 hours, the mixture is cooled andfiltered. Concentration in vacuo gave a solid which is triturated withMeOH (10 ml) to give7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (3.8 g, 85%). MS(ESI) m/z 379.2 (M+H)⁺ (method B).

¹H NMR (400 MHz, DMSO-d6): 10.47 (1H, s), 9.94 (1H, s), 8.91 (1H, s),8.82 (1H, d), 8.49 (1H, d), 8.18 (1H, dd), 6.69 (1H, s), 4.83-4.73 (1H,m), 3.06 (6H, s), 2.48-2.38 (2H, m), 2.02 (4H, s), 1.68 (2H, d).

Dess-Martin Periodinane Method

A suspension of Dess-Martin periodinane (0.435 g, 1.06 mmol) indichloromethane (5 ml) and tert-BuOH (0.1 ml) is stirred at rt. for 15minutes. To this mixture is added a solution of7-cyclopentyl-2-(5-hydroxymethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide (0.3 g, 0.79 mmol) indichloromethane:THF (5 ml:7 ml) over 5 minutes. The reaction is stirredat rt. for 1 h after which ether (50 ml) and NaOH 1M (25 ml) are added.The mixture stirred vigorously for 10 minutes and then the phases areseparated. The aqueous layer is back-extracted with ether (25 ml). Thecombined organic fractions are ished with water, brine, dried (MgSO₄)filtered and the solvent evaporated to give7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as a white solid (0.244 g, 82%).

Example 75 7-Cyclopentyl-2-[5-((3S,5R)-3,5-dimethyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (150 mg, 0.396 mmol) and tert-butyl(2R,6S)-2,6-dimethylpiperazine-1-carboxylate (93 mg, 0.436 mmol) gave(2S,6R)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2,6-dimethyl-piperazine-1-carboxylicacid tert-butyl ester (170 mg, 74%) [following SiO₂ chromatography,eluting with 0-10% MeOH)/dichloromethane]. MS(ESI) m/z 577.3 (M+H)⁺

Following General Procedure A,(2S,6R)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2,6-dimethyl-piperazine-1-carboxylicacid tert-butyl ester (170 mg, 0.295 mmol) gave7-cyclopentyl-2-[5-((3S,5R)-3,5-dimethyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (128 mg, 91%) [following SiO₂ chromatography, elutingwith 0-10% (2M NH₃ in MeOH)/DCM]. MS(ESI) m/z 477.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d₃-OD): 8.78 (1H, s), 8.43 (1H, d), 8.22 (1H, d),7.79 (1H, dd), 6.66 (1H, s), 4.83-4.75 (1H, m), 3.61 (2H, s), 3.30-3.21(2H, m), 3.18 (6H, s), 3.01 (2H, d), 2.65-2.52 (2H, m), 2.11 (4H, d),1.98 (2H, t), 1.84-1.71 (2H, m), 1.24 (6H, d).

Example 777-Cyclopentyl-2-{5-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.264 mmol) and 1-(2-methoxyethyl)piperazine(42 mg, 0.291 mmol) gave7-cyclopentyl-2-{5-[4-(2-methoxy-ethyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 71%) [following SiO₂ chromatography, elutingwith 0-10% (2M NH₃ in MeOH)/dichloromethane]. MS(ESI) m/z 507.3 (M+H)⁺(method A).

¹H NMR (400 MHz, Me-d₃-OD): 8.77 (1H, s), 8.43 (1H, d), 8.21 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.74 (1H, m), 3.61-3.47 (4H, m), 3.35(3H, s), 3.18 (6H, s), 2.81-2.41 (12H, m), 2.11 (4H, d), 1.77 (2H, d).

Example 627-Cyclopentyl-2-[5-(4-isopropyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (150 mg, 0.396 mmol) and N-isopropoylpiperazine (56mg, 0.436 mmol) gave7-cyclopentyl-2-[5-(4-isopropyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (103 mg, 53%) [following SiO₂ chromatography, elutingwith 0-10% (2M NH₃ in MeOH)/dichloromethane]. MS(ESI) m/z 491.3 (M+H)⁺(method A).

¹H NMR (400 MHz, Me-d3-OD): 8.78 (1H, s), 8.43 (1H, d), 8.22 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.84-4.77 (1H, m), 3.56 (2H, s), 3.33-3.28(1H, m), 3.18 (6H, s), 2.78-2.49 (10H, m), 2.11 (4H, d), 1.78 (2H, d),1.11 (6H, d).

Example 857-Cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (150 mg, 0.396 mmol) and N-(2-hydroxyethyl)piperazine(57 mg, 0.436 mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (78 mg, 40%) [following SiO₂chromatography, eluting with 0-10% (2M NH₃ in MeOH)/DCM]. MS(ESI) m/z493.3 (M+H)⁺ (method A).

¹H NMR (400 MHz, Me-d₃-OD): 8.77 (1H, s), 8.43 (1H, d), 8.21 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.74 (1H, m), 3.69 (2H, t), 3.55 (2H,s), 3.18 (6H, s), 2.73-2.47 (12H, m), 2.11 (4H, d), 1.78 (2H, d).

Example 347-Cyclopentyl-2-[5-(4-methanesulfonyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (150 mg, 0.396 mmol) and 1-methanesulfonyl-piperazine(72 mg, 0.436 mmol) gave7-cyclopentyl-2-[5-(4-methanesulfonyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (97 mg, 46%) [following SiO₂chromatography, eluting with 0-10% (2M NH₃ in MeOH)/DCM].

MS(ESI) m/z 527.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, Me-d₃-OD): 8.77 (1H, s), 8.43 (1H, d), 8.22 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.80-4.75 (1H, m), 3.60 (2H, s), 3.30-3.25(4H, m), 3.18 (6H, s), 2.86 (3H, s), 2.66-2.54 (6H, m), 2.11 (4H, d),1.83-1.72 (2H, m).

Example 612-[5-(4-Acetyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (200 mg, 0.528 mmol) and 1-acetyl piperazine (75 mg,0.581 mmol) gave2-[5-(4-acetyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as an off white solid (96 mg, 37%) [following SiO₂chromatography, eluting with 0-10% (2M NH₃ in MeOH)/dichloromethane].MS(ESI) m/z 491.2 (M+H)⁺ (method A).

¹H NMR (400 MHz, Me-d₃-OD): 8.77 (1H, s), 8.43 (1H, d), 8.22 (1H, d),7.81 (1H, dd), 6.65 (1H, s), 4.84-4.74 (1H, m), 3.63 (2H, t), 3.57 (4H,d), 3.18 (6H, s), 2.65-2.44 (6H, m), 2.19-2.03 (7H, m), 1.78 (2H, d).

Example 547-Cyclopentyl-2-[5-(3-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Following General Procedure C,7-Cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (0.298 g, 0.788 mmol) and2-methyl-piperazine-1-carboxylic acid tert-butyl ester (0.316 g, 1.58mmol) gave 4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester a yellowish oil (0.341 g, 77%) [followingpurification by SiO₂ chromatography eluting with 2-6% MeOH/DCM].

MS(ESI) m/z 563.3 (M+H)⁺

Following General Procedure A,4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (0.341 g, 0.606 mmol) gave7-cyclopentyl-2-[5-(3-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide as a white solid (70 mg, 25%)[following SiO₂ chromatography, eluting with 0-10% (2M NH₃ inMeOH)/dichloromethane).

MS(ESI) m/z 463.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.77 (1H, s), 8.42 (1H, d), 8.21 (1H, s),7.80 (1H, dd), 6.65 (1H, s), 4.82-4.73 (1H, m), 3.54 (2H, s), 3.18 (6H,s), 3.04-2.94 (1H, m), 2.94-2.76 (4H, m), 2.67-2.51 (2H, m), 2.20-2.01(5H, m), 1.85-1.73 (3H, m), 1.08 (3H, d).

Example 10A7-Cyclopentyl-2-[5-(3-hydroxy-azetidin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide and 3-hydroxy-azetidinium chloride (73 mg, 0.667mmol) gave7-cyclopentyl-2-[5-(3-hydroxy-azetidin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide as a light yellow solid(0.131 g, 47%) [following SiO₂ chromatography, eluting with 0-5% (2M NH₃in MeOH)/DCM). MS(ESI) m/z 436.2 (M+H)⁺ (method B).

¹H NMR (400 MHz, CDCl₃): 8.78 (1H, s), 8.59-8.41 (2H, m), 8.23 (1H, d),7.65 (1H, dd), 6.47 (1H, s), 4.88-4.72 (1H, m), 4.55-4.42 (1H, m),3.84-3.42 (4H, m), 3.17 (6H, s), 3.02 (2H, t), 2.68-2.53 (2H, m), 2.07(4H, d), 1.73 (2H, d).

Example 667-Cyclopentyl-2-{5-[((R)-2-hydroxy-1-methyl-ethylamino)-methyl]-pyridin-2lamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Following General Procedure C,7-Cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 0.132 mmol) and (R)-2-amino propan-1-ol (20mg, 0.264 mmol) gave7-cyclopentyl-2-{5-[((R)-2-hydroxy-1-methyl-ethylamino)-methyl]-pyridin-2-yl-amino-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide as a white solid (36 mg,62%). MS(ESI) m/z 438.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, CDCl3): 8.75 (1H, s), 8.47 (1H, d), 8.28 (1H, d), 7.73(1H, dd), 6.47 (1H, s), 4.88-4.77 (1H, m), 3.90 (1H, d), 3.77 (1H, d),3.67 (1H, dd), 3.36 (1H, dd), 3.18 (6H, s), 2.98-2.88 (1H, m), 2.67-2.53(2H, m), 2.10 (4H, d), 1.75 (2H, d), 1.16 (3H, d).

Example 557-Cyclopentyl-2-[5-((S)-3-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure C,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (3.8 g, 10.05 mmol) and(S)-2-methyl-piperazine-1-carboxylic acid tert-butyl ester (5.03 g,25.13 mmol) gave(S)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester as a white solid (2.45 g, 45%) [following SiO₂chromatography, eluting with 50% EtOAc/petroleum ether). MS(ESI) m/z563.3 (M+H)⁺

Following General Procedure A,(S)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (2.45 g, 4.35 mmol) gave7-cyclopentyl-2-[5-((S)-3-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (1.3 g, 65%) [following SiO₂ chromatography, elutingwith 5% (NH₃ 2.0 M in MeOH)/dichloromethane). MS(ESI) m/z 463.3 (M+H)⁺(method A).

¹H NMR (400 MHz, Me-d3-OD): 8.77 (1H, s), 8.42 (1H, d), 8.21 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.54 (2H, s), 3.18 (6H,s), 3.03-2.92 (1H, m), 2.92-2.77 (4H, m), 2.67-2.51 (2H, m), 2.20-2.01(5H, m), 1.86-1.70 (3H, m), 1.07 (3H, d).

Example 767-Cyclopentyl-2-[5-(4-hydroxy-piperidin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (0.2 g, 0.529 mmol) and piperidin-4-ol (56 mg, 0.556mmol) gave7-cyclopentyl-2-[5-(4-hydroxy-piperidin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide is obtained as a white solid (25 mg, 10%) [followingpurification by preparative LCMS and further purification by SiO₂chromatography, eluting with 0-7% (2N NH₃ in MeOH/EtOAc). MS(ESI) m/z464.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.77 (1H, s), 8.42 (1H, d), 8.20 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.72-3.60 (1H, m), 3.54(2H, s), 3.18 (6H, s), 2.83 (2H, d), 2.67-2.50 (2H, m), 2.24 (2H, t),2.11 (4H, d), 1.89 (2H, d), 1.77 (2H, d), 1.69-1.51 (2H, m).

Example 427-Cyclopentyl-2-(5-piperazin-1-ylmethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyri-midine-6-carboxylic acid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (0.35 g, 0.926 mmol) and piperazine-1-carboxylic acidtert-butyl ester (0.19 g, 1.02 mmol) gave4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester as a white solid (0.333 g, 60%). The material isused directly in the next step. MS(ESI) m/z 549.3 (M+H)⁺

Following General Procedure A,4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-piperazine-1-carboxylicacid tert-butyl ester (0.333 g, 0.607 mmol) gave7-cyclopentyl-2-(5-piperazin-1-ylmethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyri-midine-6-carboxylic acid dimethylamide as a white powder (135 mg,50%) [following treatment with DOWEX 550A and further purification bySiO₂ chromatography, eluting with 0-12% (2N NH₃ inMeOH/dichloromethane). MS(ESI) m/z 449.4 (M+H)⁺ (method D).

¹H NMR (400 MHz, Me-d3-OD): 8.78 (1H, s), 8.43 (1H, d), 8.21 (1H, d),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.54 (2H, s), 3.18 (6H,s), 2.88 (4H, t), 2.79-2.35 (6H, m), 2.11 (4H, d), 1.78 (2H, d).

Example 437-Cyclopentyl-2-[5-(4-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (0.2 g, 0.529 mmol) and N-methyl piperazine (58 mg,0.582 mmol) gave7-cyclopentyl-2-[5-(4-methyl-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as a white off solid (0.144 g, 60%) [followingfurther purification by SiO₂ chromatography, eluting with 2-5% (2N NH₃in MeOH/dichloromethane]. MS(ESI) m/z 463.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.78 (1H, s), 8.43 (1H, d), 8.21 (1H, s),7.79 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.56 (2H, s), 3.18 (6H,s), 2.98-2.35 (10H, m), 2.30 (3H, s), 2.11 (4H, d), 1.78 (2H, d).

Example 737-Cyclopentyl-2-[5-(3,8-diaza-bicyclo[3.2.1]oct-3-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure B,7-cyclopentyl-2-(5-formyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (0.153 g, 0.405 mmol) and3,8-diaza-bicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (95mg, 0.445 mmol) gave3-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-3,8-diaza-bicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester as an off white solid (0.186 g, 80%) [followingpurification by SiO₂ chromatography, eluting with 0-5% MeOH/EtOAc].MS(ESI) m/z 575.3 (M+H)⁺

Following General Procedure A,3-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-ylmethyl]-3,8-diaza-bicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester (0.186 g, 0.324 mmol) gave7-cyclopentyl-2-[5-(3,8-diaza-bicyclo[3.2.1]oct-3-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as a white solid (0.105 g, 68%) [followingpurification by SiO₂ chromatography, eluting with 0-5% (2M NH₃ inMeOH)/dichloromethane]. MS(ESI) m/z 475.3 (M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.77 (1H, s), 8.38 (1H, d), 8.18 (1H, s),7.76 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.49 (2H, s), 3.42 (2H,s), 3.18 (6H, s), 2.68 (2H, dd), 2.64-2.50 (2H, m), 2.29 (2H, d),2.23-2.01 (4H, m), 2.01-1.85 (2H, m), 1.77 (4H, d).

Example 652-{5-[4-(2-Amino-acetyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

By repeating procedures outlined in Example 12,7-cyclopentyl-2-(5-piperazin-1-ylmethyl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyri-midine-6-carboxylic acid dimethylamide (0.108 g, 0.241 mmol) andtert-butoxycarbonylamino-acetic acid (42 mg, 0.241 mmol) gave2-{5-[4-(2-amino-acetyl)-piperazin-1-ylmethyl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxy-licacid dimethylamide as a white solid (87 mg, 69%). MS(ESI) m/z 506.3(M+H)⁺ (method B).

¹H NMR (400 MHz, Me-d3-OD): 8.78 (1H, s), 8.43 (1H, d), 8.22 (1H, s),7.80 (1H, dd), 6.65 (1H, s), 4.83-4.72 (1H, m), 3.71-3.62 (2H, m), 3.58(2H, s), 3.54-3.39 (4H, m), 3.18 (6H, s), 2.67-2.55 (2H, m), 2.55-2.41(4H, m), 2.20-2.03 (4H, m), 1.86-1.68 (2H, m).

Example 607-Cyclopentyl-2-[5-(4-methyl-3-oxo-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Method B, 6-chloro-pyridine-3-carbaldehyde (500 mg,3.532 mmol) and 1-methylpiperazine-2-one hydrochloride (559 mg, 3.709mmol) gave 4-(6-chloro-pyridin-3-ylmethyl)-1-methyl-piperazin-2-one (712mg, 84%) [following SiO₂ chromatography, eluting with 0-10%MeOH/dichloromethane]. MS(ESI) m/z 240.1 (M+H)⁺

By repeating procedures outlined in Example C step 2 (except thatbenzhydrylidene intermediate is extracted using EtOAc),4-(6-chloro-pyridin-3-ylmethyl)-1-methyl-piperazin-2-one (712 mg, 2.970mmol) gave 4-(6-amino-pyridin-3-ylmethyl)-1-methyl-piperazin-2-one (31mg, 28%) [following SiO₂ chromatography, eluting with 0-10% (2M NH₃ inMeOH)/dichloromethane]. MS(ESI) m/z 221.3 (M+H)⁺

Following Buchwald method B,4-(6-amino-pyridin-3-ylmethyl)-1-methyl-piperazin-2-one (30 mg, 0.136mmol) and2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (33 mg, 0.113 mmol) give7-cyclopentyl-2-[5-(4-methyl-3-oxo-piperazin-1-ylmethyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (11 mg, 21%) [following SiO₂ chromatography elutingwith 1-10% (2M NH₃ in MeOH)/DCM] MS(ESI) m/z 477.3 (M+H)⁺ (method B)

¹H NMR (400 MHz, Me-d₃-OD): 8.78 (1H, s), 8.44 (1H, d), 8.23 (1H, d),7.81 (1H, dd), 6.65 (1H, s), 4.83-4.76 (1H, m), 3.62 (2H, s), 3.40 (2H,t), 3.19-3.15 (8H, m), 2.97 (3H, s), 2.78 (2H, t), 2.64-2.53 (2H, m),2.11 (4H, d), 1.77 (2H, d).

Dimethylated Amide Series General Procedure F2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide

In a large sealed tube is added 5-bromo-2,4-dichloropyrimidine (3 g,13.2 mmol) in 100 mL of EtOH. Then cyclopentyl amine (1.95 mL, 19.75mmol) and N,N′-diisopropylethylamine (3.36 mL, 19.8 mmol) are added tothe solution at rt. The solution is then stirred rt overnight. Solventis evaporated and the crude is purified using silica gel chromatography(15% ethyl acetate/85% hexane) to give(5-bromo-2-chloro-pyrimidin-4-yl)-cyclopentyl-amine as a white solid(3.25 g, 89%). MS(ESI) m/z 278.4 (M+H)⁺

A mixture of (5-bromo-2-chloro-pyrimidin-4-yl)-cyclopentyl amine (1 g,3.6 mmol), propiolaldehydediethylacetel (550 mg, 4.3 mmol), PdCl₂(PPh₃)₂(252 mg, 0.36 mmol), CuI (70 mg, 0.36 mmol), 20 mL of Et₃N and 5 mL ofDMF is degassed and heated to 100° C. After 13 h, solvent are removedand column is run using 5% ethyl acetate in heptane to 10% ethyl acetatein heptane. Product concentrated to give[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-cyclopentyl amine(500 mg, 43%). MS(ESI) m/z 324.5 (M+H)⁺

A mixture of[2-chloro-5-(3,3-diethoxy-prop-1-ynyl)-pyrimidin-4-yl]-cyclopentyl amine(5.21 g, 16 mmol) in THF is added 1M tetra-n-butylammonium fluoride inTHF (TBAF) (97 mL, 97 mmol) and heated to 65° C. for 2 hour. Solvent isremoved and column is run using heptane/ethyl acetate from 5% to 15% togive 2-chloro-7-cyclopentyl-6-diethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine(4.26 g, 82%). MS(ESI) m/z 324.5 (M+H)⁺

A mixture of2-chloro-7-cyclopentyl-6-diethoxymethyl-7H-pyrrolo[2,3-d]pyrimidine(4.26 g, 13 mmol) in dioxane is added concentrated HCl. After reactionis completed within 10 min, water is added and then extracted with ethylacetate. Solvent is removed to give a brown color crude product. Columnis run using heptane/ethyl acetate (6:4) to give2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde (2.69g, 82%). MS(ESI) m/z 350.4 (M+H)⁺

A mixture of2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde (2.69g, 11 mmol) in DMF is added oxone (7.2 g, 12 mmol) and stirred for 6 h.After the reaction is complete, water is added and a yellow solid isprecipitated to give2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid(2.69 g, 85%). MS(ESI) m/z 266.4 (M+H)⁺

2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid(1.07 g, 4.03 mmol), HBTU (1.53 g, 4.03 mmol) and diisopropylethylamine(2 mL, 12.1 mmol) are dissolves in dimethylformamide (20 mL). 2 Msolution of dimethylamine in ethanol (2.4 mL, 4.8 mmol) is added and themixture is stirred for 30 minutes to achieve complete conversion. Thereaction mixture is diluted with ethyl acetate and washed with saturatedaqueous sodium hydrogen carbonate, water, then brine. The organic phaseis dried (Na₂SO₄), filtered and concentrated. Purification bychromatography on silicagel (ethyl acetate:heptane) provides2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (927 mg, 79% yield) MS(ESI) m/z 293.1 (M+H)⁺

Example 17-Cyclopentyl-2-(pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (100 mg, 0.34 mmol) and pyridine-2-ylamine (64 mg, 0.68mmol) gave7-cyclopentyl-2-(pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (350 mg, 84%).

MS(ESI) m/z 351.1 (M+H)⁺

Example 747-Cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B, then General Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (300 mg, 1.02 mmol) and 5-piperazin-1-yl-pyridin-2-ylamine(314 mg, 1.13 mmol) gave7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (142 mg, 36%). MS(ESI) m/z 435.3 (M+H)⁺

Alkylated Analogues General Procedure D Example 787-Cyclopentyl-2-[5-(4-ethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) in 20 mL of THF is addedpotassium carbonate (100 mg, 0.689 mmol) then bromoethane (75 mg, 0.687mmol). The reaction mixture is heated at 70° C. for 18 h. Following SiO₂chromatography, eluting with 0-10% (2M NH₃ in MeOH)/dichloromethan] gave7-cyclopentyl-2-[5-(4-ethyl-piperazin-1-yl)-pyridin-2-ylamino]-7Hpyrrolo[2,3d]pyrimidine-6-carboxylic acid dimethylamide (67 mg, 63%).MS(ESI) m/z 463.3 (M+H)⁺

Example 867-Cyclopentyl-2-{5-[4-(2-fluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) and 1-bromo-2-fluoroethane (88mg, 0.687 mmol) gave7-cyclopentyl-2-{5-[4-(2-fluoro-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (51 mg, 80%). MS(ESI) m/z 481.3 (M+H)⁺

Example 267-Cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (34 mg, 0.072 mmol) and 2-bromo ethanol (9 mg, 0.216mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (12 mg, 32%). MS(ESI) m/z 479.3 (M+H)⁺

Example 957-Cyclopentyl-2-{5-[4-(2-isopropoxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) and 2-(2-bromoethoxy)propane(200 mg, 0.252 mmol) gave7-cyclopentyl-2-{5-[4-(2-isopropoxy-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (103 mg, 86%). MS(ESI) m/z 521.3 (M+H)⁺

General Procedure E Example 577-Cyclopentyl-2-{5-[4-((R)-2-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (55 mg, 0.123 mmol) and (R)-2-methyl-oxirane (250 mg,4.3 mmol) in 5 mL of ethanol is heated at 70° C. for 18 h. FollowingSiO₂ chromatography, eluting with 0-10% (2M NH₃ inMeOH)/dichloromethane] gave7-cyclopentyl-2-{5-[4-((R)-2-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (10 mg, 16%). MS(ESI) m/z 493.3 (M+H)⁺

¹H NMR (400 MHz, CDCl₃): 8.70 (1H, s), 8.32 (1H, d), 7.96 (1H, s), 7.80(1H, s), 7.28 (1H, d), 6.45 (1H, s), 5.32 (1H, s), 4.86-4.77 (1H, s),3.85 (2H, t), 3.44 (2H, t), 3.18 (6H, s), 2.98 (3H, s), 2.62-2.59 (2H,m), 2.11-2.02 (3H, m); 1.74-1.63 (3H, m).

Example 567-Cyclopentyl-2-{5-[4-((S)-2-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure E,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (48 mg, 0.110 mmol) and (S)-2-methyl oxirane (121 mg,0.22 mmol) gave7-cyclopentyl-2-{5-[4-((S)-2-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (10 mg, 16%). MS(ESI) m/z 493.3 (M+H)⁺

Example 717-Cyclopentyl-2-{5-[4-(2-hydroxy-2-methyl-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure E,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 0.115 mmol) and 2,2-dimethyloxirane (72 mg,0.805 mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-2-methyl-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (17 mg, 29%). MS(ESI) m/z 507.3 (M+H)⁺

Example 217-Cyclopentyl-2-{5-[4-(3-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) and 3-bromo propan-1-ol (80 mg,0.574 mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-2-methylpropyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (55 mg, 50%). MS(ESI) m/z 493.3 (M+H)⁺

Example 447-Cyclopentyl-2-{5-[4-(3-hydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) and 3-bromo propane-1,2-diol(106 mg, 0.687 mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-2-methyl-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (29 mg, 24%). MS(ESI) m/z 509.3 (M+H)⁺

Example 467-Cyclopentyl-2-{5-[4-((R)-2,3-dihydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and R-(+) glycidol (51 mg, 0.691mmol) gave7-cyclopentyl-2-{5-[4-((R)-2,3-dihydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (56 mg, 47%). MS(ESI) m/z 509.3 (M+H)⁺

Example 297-Cyclopentyl-2-{5-[4-((S)-2,3-dihydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure E,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and (S)-(+) glycidol (51 mg,0.691 mmol) gave7-cyclopentyl-2-{5-[4-((S)-2,3-dihydroxy-propyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (60 mg, 50%). MS(ESI) m/z 509.3 (M+H)⁺

Example 797-Cyclopentyl-2-[5-(4-cyclopentyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and bromo cyclopentane (103 mg,0.691 mmol) gave7-cyclopentyl-2-[5-(4-cyclopentyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (85 mg, 71%). MS(ESI) m/z 503.3 (M+H)⁺

Example 637-Cyclopentyl-2-[5-(4-isopropyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (30 mg, 0.069 mmol) in 10 mL of dichloromethane isadded 1 mL of acetone and NaB(OAc)₃H (30 mg, 0.138 mmol). The resultingmixture is stirred at room temperature for 18 h. Following purificationby preparative LCMS gave7-cyclopentyl-2-[5-(4-isopropyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (20 mg, 61%). MS(ESI) m/z 477.3 (M+H)⁺

Example 367-Cyclopentyl-2-{5-[4-(2-hydroxy-1-methyl-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 2-bromo propan-1-ol (96 mg,0.690 mmol) gave7-cyclopentyl-2-{5-[4-(2-hydroxy-1-methyl-ethyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (28 mg, 25%). MS(ESI) m/z 493.4 (M+H)⁺

Example 1012-{4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionicacid methyl ester

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 2-bromo propionic acidmethyl ester (31 mL, 0.28 mmol) gave2-{4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionicacid methyl ester (46 mg, 39%). MS(ESI) m/z 521.4 (M+H)⁺

Example 1032-{4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionicacid

To a solution of2-{4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionicacid methyl ester (250 mg, 0.48 mmol) in 10 mL of THF is added asolution of LiOH (19 mg, 48 mmol) in 10 mL of H₂O. After 18 h stirringat room temperature, the resulting mixture is concentrated and dilutedwith H₂O and adjusted to pH=6 with 1 N HCl. Ished with dichloromethanethen solids precipitated and collected to give2-{4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-piperazin-1-yl}-propionicacid (225 mg, 94%). MS(ESI) m/z 507.3 (M+H)⁺

Example 692-[5-(4-Cyclohexylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and bromomethyl cyclohexane (122mg, 0.690 mmol) gave2-[5-(4-cyclohexylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (75 mg, 63%). MS(ESI) m/z 531.4 (M+H)⁺

Example 927-Cyclopentyl-2-[5-(4-isobutyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 1-bromo-2-methyl propane (94mg, 0.690 mmol) gave7-cyclopentyl-2-[5-(4-isobutyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (45 mg, 41%). MS(ESI) m/z 491.3 (M+H)⁺

Example 997-Cyclopentyl-2-{5-[4-(2-methyl-butyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 1-bromo-2-methyl butane (103mg, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(2-methyl-butyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 42%). MS(ESI) m/z 505.3 (M+H)⁺

Example 687-Cyclopentyl-2-{5-[4-(4-methyl-pentyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 1-bromo-4-methyl pentane(103 mg, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(4-methyl-pentyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 42%). MS(ESI) m/z 519.4 (M+H)⁺

Example 102-[5-(4-Carbamoylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 2-bromo acetamide (95 mg,0.690 mmol) gave 2-[5-(4-carbamoylmethylpiperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 45%). MS(ESI) m/z 492.4 (M+H)⁺

Example 7 2-[5-(4-Acetylpiperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (30 mg, 0.230 mmol) in 5 mL of dichloromethane. Added0.5 mL of acetic anhydride. After 10 min, reaction is complete andtrituration with acetonitrile gave2-[5-(4-acetyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (30 mg, 91%). MS(ESI) m/z 477.3 (M+H)⁺

General Procedure G Example 277-Cyclopentyl-2-[5-(4-cyclopropanecarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and cyclopropanecarbonylchloride (22 mL, 0.690 mmol) in 5 mL of CH₂Cl₂ is added a solution ofEt₃N (64 mL, 0.459 mmol) and stirred at rt for 18 h. The resultingmixture is concentrated and diluted with saturated NaHCO₃ and extractedwith ethyl acetate (3×100 mL). The combine organics are dried overNa₂CO₃ and preparative HPLC to give2-[5-(4-carbamoylmethyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (81 mg, 68%). MS(ESI) m/z 503.3 (M+H)⁺

Example 232-[5-(4-Cyclohexanecarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and cyclohexane carbonylchloride (37 mg, 0.690 mmol) gave2-[5-(4-cyclohexanecarbonyl-piperazin-1-yl)-pyridin-2-ylamino]-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (63 mg, 49%). MS(ESI) m/z 545.3 (M+H)⁺

Example 902-{5-[4-(2-Cyclohexyl-acetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and cyclohexyl acetyl chloride(39 mL, 0.690 mmol) gave 2-{5-[4-(2-cyclohexylacetyl)-piperazin-1-yl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (61 mg, 47%). MS(ESI) m/z 559.4 (M+H)⁺

Example 917-Cyclopentyl-2-{5-[4-(3-cyclopentyl-propionyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and 3-cyclopentyl propionylchloride (39 mL, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(3-cyclopentyl-propionyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (57 mg, 44%). MS(ESI) m/z 559.4 (M+H)⁺

Example 227-Cyclopentyl-2-{5-[4-(pyrrolidine-1-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and pyrrolidine-1-carbonylchloride (25 mL, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(pyrrolidine-1-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (84 mg, 70%). MS(ESI) m/z 532.3 (M+H)⁺

Example 947-Cyclopentyl-2-{5-[4-(piperidine-1-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and piperidine-1-carbonylbromide (32 mL, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(piperidine-1-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (83 mg, 64%). MS(ESI) m/z 546.3 (M+H)⁺

Example 387-Cyclopentyl-2-{5-[4-(morpholine-4-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure G,7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.230 mmol) and morpholine-4-carbonylchloride (38 mg, 0.690 mmol) gave7-cyclopentyl-2-{5-[4-(morpholine-4-carbonyl)-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (80 mg, 62%). MS(ESI) m/z 548.3 (M+H)⁺

Example 30(R)-4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (200 mg, 0.200 mmol) and(R)-2-methyl-piperazine-1-carboxylic acid tert-butyl ester (200 mg,0.682 mmol) gave(R)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (131 mg, 35%). MS(ESI) m/z 549.5 (M+H)⁺

Example 31(S)-4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (200 mg, 0.200 mmol) and(S)-2-methyl-piperazine-1-carboxylic acid tert-butyl ester (200 mg,0.682 mmol) gave(S)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (157 mg, 42%). MS(ESI) m/z 549.5 (M+H)⁺

Example 167-Cyclopentyl-2-[5-((R)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure A,(R)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (131 mg, 0.200 mmol) gave7-cyclopentyl-2-[5-((R)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (55 mg, 50%). MS(ESI) m/z 449.3 (M+H)⁺

¹H NMR (400 MHz, CDCl₃): 8.71 (1H, s), 8.38 (1H, d), 8.03 (1H, s), 7.80(1H, s), 7.36 (1H, d), 6.46 (1H, s), 4.84-4.80 (1H, m), 3.46 (3H, d),3.18 (6H, s), 3.14-3.05 (2H, m), 2.82-2.75 (1H, m), 2.60-2.55 (3H, m),2.47-2.41 (1H, m), 2.10-2.04 (4H, m), 1.94-1.67 (4H, m).

Example 817-Cyclopentyl-2-{5-[(R)-4-(2-hydroxy-ethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-[5-((R)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (43 mg, 0.095 mmol) and 2-bromo ethanol (13 mg, 0.105mmol) gave7-cyclopentyl-2-{5-[(R)-4-(2-hydroxy-ethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (32 mg, 68%). MS(ESI) m/z 493.3 (M+H)⁺

Example 177-Cyclopentyl-2-[5-((S)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure A,(S)-4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2-methyl-piperazine-1-carboxylicacid tert-butyl ester (145 mg, 0.200 mmol) gave7-cyclopentyl-2-[5-((S)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

(86 mg, 72%). MS(ESI) m/z 449.3 (M+H)⁺

Example 827-Cyclopentyl-2-{5-[(S)-4-(2-hydroxy-ethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-[5-((S)-3-methyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (74 mg, 0.17 mmol) and 2-bromo ethanol (23 mg, 0.18mmol) gave7-cyclopentyl-2-{5-[(S)-4-(2-hydroxy-ethyl)-3-methyl-piperazin-1-yl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (34 mg, 42%). MS(ESI) m/z 493.3 (M+H)⁺

Example 727-Cyclopentyl-2-[5-(3,3-dimethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (50 mg, 0.17 mmol) and4-(6-amino-pyridin-3-yl)-2,2-dimethyl-piperazine-1-carboxylic acidtert-butyl ester (58 mg, 0.15 mmol) gave7-cyclopentyl-2-[5-(3,3-dimethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (20 mg, 25%). MS(ESI) m/z 463.3 (M+H)⁺

Example 247-Cyclopentyl-2-[5-(3,5-dimethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure A,4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-2,6-dimethyl-piperazine-1-carboxylicacid tert-butyl ester (150 mg, 0.27 mmol) gave7-cyclopentyl-2-[5-(3,5-dimethyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (70 mg, 58%). MS(ESI) m/z 463.3 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.33 (1H, s), 8.76 (1H, s), 8.15 (1H, d),7.98 (1H, s), 7.43, (1H, d), 6.61 (1H, s), 4.76-4.72 (1H, m), 3.50-3.48(2H, m), 3.08-3.05 (3H, m), 2.89-2.86 (2H, m), 2.50 (12H, s), 2.48-2.43(2H, m), 2.14-2.05 (2H, m), 2.00-1.90 (2H, m), 1.70-1.60 (1H, m).

Example 47-Cyclopentyl-2-[5-(3-oxo-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (100 mg, 0.34 mmol) and4-(6-amino-pyridin-3-yl)-piperazin-2-one (111 mg, 0.578 mmol) gave7-cyclopentyl-2-[5-(3-oxo-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (35 mg, 35%). MS(ESI) m/z 449.2 (M+H)⁺

Example 397-Cyclopentyl-2-[5-((S)-3-hydroxy-pyrrolidin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (101 mg, 0.35 mmol) andN-{(E)-2-[(S)-3-(tert-butyl-dimethyl-silanyloxy)-pyrrolidin-1-yl]-vinyl}-acrylamidine(153 mg, 0.52 mmol), followed by deprotection with TBAF to give7-cyclopentyl-2-[5-((S)-3-hydroxy-pyrrolidin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (98 mg, 65%). MS(ESI) m/z 436.3 (M+H)⁺

Example 327-Cyclopentyl-2-[5-(3-hydroxy-azetidin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of 5-bromo-2-niropyridine (0.54 g, 2.66 mmol),azetidin-3-ol hydrochloride (0.46 g, 4.17 mmol) and tetrabutylammoniumiodide (0.103 g, 0.278 mmol) in 6 mL of DMSO is added potassiumcarbonate (1.06 g, 7.68 mmol). The resulting mixture is heated to 80° C.for 3 h. Poured into ethyl acetate/NaHCO₃ solution. Extracted with ethylacetate (2×250 mL). The organic layer is ished with brine and dried overNa₂SO₄. Concentrated to give 1-(6-nitro-pyridin-3-yl)-azetidin-3-ol (153mg, 29%). MS(ESI) m/z 240.1 (M+H)⁺

To a solution of 1-(6-nitro-pyridin-3-yl)-azetidin-3-ol (154 mg, 0.779mmol) in 2 mL of DMF is added Et₃N (0.2 mL, 0.15 mmol), TBDMSCl (117 mg,0.776 mmol). The resulting mixture is stirred at room temperature for 2h. Poured into EtOAc/NaHCO₃. the aqueous layer is extracted with ethylacetate (2×50 mL). The combined organic layers are ished with brine,dried over Na₂SO₄. Concentrated to give5-[3-(tert-butyl-dimethyl-silanyloxy)-azetidin-1-yl]-2-nitro-pyridine(175 mg, 73%).

To a suspension of5-[3-(tert-butyl-dimethyl-silanyloxy)-azetidin-1-yl]-2-nitro-pyridine(124 mg, 0.401 mmol) in 5 mL of ethanol is added iron powder (206 mg,3.68 mmol) then 2 mL of NH₄Cl solution. The resulting mixture is heatedat 80° C. for 3 h and filtered through celite and concentrated. Theresulting dark solid is divided between ethyl acetate and water. Theaqueous phase is extracted with ethyl acetate (2×50 mL). The combineorganic layers are ished with brine, dried over Na₂SO₄ and concentratedto give5-[3-(tert-butyl-dimethyl-silanyloxy)-azetidin-1-yl]-pyridin-2-ylamine(105 mg, 94%).

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (120 mg, 0.411 mmol) andN-{(E)-2-[(S)-3-(tert-butyl-dimethyl-silanyloxy)-pyrrolidin-1-yl]-vinyl}-acrylamidine(112 mg, 0.401 mmol), followed by deprotection with 2 mL of TBAF to give7-cyclopentyl-2-[5-(3-hydroxy-azetidin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (18 mg, 46%). MS(ESI) m/z 422.5 (M+H)⁺

Example 592-{5-[(2-Amino-ethyl)-methyl-amino]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (200 mg, 0.68 mmol) and{2-[(6-amino-pyridin-3-yl)-methyl-amino]-ethyl}-carbamic acid tert-butylester (200 mg, 0.75 mmol), followed by deprotection using GeneralProcedure A to give2-{5-[(2-amino-ethyl)-methyl-amino]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 34%). MS(ESI) m/z 423.4 (M+H)⁺

Example 837-Cyclopentyl-2-{5-[(2-hydroxy-ethyl)-methyl-amino]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (25 mg, 0.85 mmol) and [2-(tert-butyl dimethylsilanyloxy)-ethyl]-methyl amine (27 mg, 0.094 mmol), followed bydeprotection using 0.6 mL of TBAF in 2 mL of THF to give7-cyclopentyl-2-{5-[(2-hydroxy-ethyl)-methyl-amino]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (18 mg, 72%). MS(ESI) m/z 424.2 (M+H)⁺

¹H NMR (400 MHz, CDCl₃): 8.78 (1H, s), 8.59-8.41 (2H, m), 8.23 (1H, d),7.65 (1H, dd), 6.47 (1H, s), 4.88-4.72 (1H, m), 4.55-4.42 (1H, m),3.84-3.42 (4H, m), 3.17 (6H, s), 3.02 (2H, t), 2.68-2.53 (2H, m), 2.07(4H, d), 1.73 (2H, d).

Example 37-Cyclopentyl-2-[5-(piperidin-4-ylcarbamoyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (25 mg, 0.85 mmol) and4-[(6-amino-pyridine-3-carbonyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester (27 mg, 0.094 mmol), followed by deprotection using 0.6mL of TBAF in 2 mL of THF to give7-cyclopentyl-2-[5-(piperidin-4-ylcarbamoyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (18 mg, 72%). MS(ESI) m/z 424.2 (M+H)⁺

Example 537-Cyclopentyl-2-[5-(methyl-piperidin-4-yl-carbamoyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (170 mg, 0.58 mmol) and4-[(6-amino-pyridine-3-carbonyl)-methyl-amino]-piperidine-1-carboxylicacid tert-butyl ester (292 mg, 0.87 mmol), followed by deprotectionusing General Procedure A to give7-cyclopentyl-2-[5-(methyl-piperidin-4-yl-carbamoyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (46 mg, 16%). MS(ESI) m/z 491.3 (M+H)⁺

Example 497-Cyclopentyl-2-[5-(piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (205 mg, 0.7 mmol) and4-(6-amino-pyridine-3-carbonyl)-piperazine-1-carboxylic acid tert-butylester (236 mg, 0.8 mmol), followed by deprotection using GeneralProcedure A to give7-cyclopentyl-2-[5-(piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (13 mg, 41%). MS(ESI) m/z 463.3 (M+H)⁺

Example 967-Cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazine-1-carbonyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-[5-(piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (200 mL, 0.43 mmol) and 2-bromo ethanol (37 mg, 0.52mmol) to give7-cyclopentyl-2-{5-[4-(2-hydroxy-ethyl)-piperazine-1-carbonyl]-pyridin-2-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 48%). MS(ESI) m/z 478.3 (M+H)⁺

Example D6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-nicotinicacid

Following General Procedure D,7-cyclopentyl-2-[5-(piperazine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (2 g, 6.83 mmol) and 6-amino-nicotinic acid methylester (1.15 g, 7.51 mmol). Followed by treatment with LiOH (1 g, 25mmol) in 320 mL of THF/H₂O gave6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)nicotinic acid (1.2 g, 55%). MS(ESI) m/z 395.3 (M+H)⁺

Example 507-Cyclopentyl-2-[5-(4-dimethylamino-piperidine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-nicotinicacid (100 mg, 0.25 mmol) (Example D) in 3 mL of DMF is addeddimethyl-piperidin-4-yl-amine (33 mg, 0.25 mmol), HBTU (140 mg, 0.38mmol), and DIPEA (0.088 mL, 0.51 mmol). After 48 h stirring at roomtemperature, the resulting mixture is concentrated and diluted withsaturated NaHCO₃ and extracted with ethyl acetate (3×100 mL). Thecombine organics are dried over Na₂CO₃ and concentrated to give areddish residue. Preparative HPLC to give7-cyclopentyl-2-[5-(4-dimethylamino-piperidine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (60 mg, 46%). MS(ESI) m/z 505.5 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.97 (1H, s), 8.85 (1H, s), 8.34 (1H, d),7.83 (1H, d), 6.65 (1H, s), 4.80-4.72 (1H, m), 4.06-4.00 (1H, s), 3.06(6H, s), 2.48-2.40 (2H, m), 2.39-2.30 (2H, m), 2.18 (6H, s), 2.05-1.95(5H, m), 1.82-1.70 (2 h, m), 1.69-1.60 (2H, m), 1.41-1.32 (2H, m),1.19-1.16 (2 h, m).

Example 877-Cyclopentyl-2-[5-(4-hydroxy-piperidine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-nicotinicacid methyl ester (200 mg, 0.49 mmol) and piperidin-4-ol (500 mg, 4.9mmol) in 5 mL of CH₂Cl₂ is added dropwise a solution of iPrMgCl (2.45mL, 4.9 mmol) at 0° C. and allow to warm up to room temperatureovernight. After 18 h, added another 10 equivalents of i-PrMgCl andstirred for another 5 h. The reaction mixture is quenched with saturatedNH₄Cl and extracted with dichloromethane (3×100 mL). The combine organicished with NaCl and dried over Na₂SO₄ and concentrated. Following SiO₂chromatography, eluting with 85/15% (CH₂Cl₂/MeOH) gave7-cyclopentyl-2-[5-(4-hydroxy-piperidine-1-carbonyl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (120 mg, 51%). MS(ESI) m/z 478.3 (M+H)⁺

Example 412-{5-[(2-Amino-ethyl)-methyl-carbamoyl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

A solution of6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-nicotinicacid (100 mg, 0.25 mmol) in DMF is added (2-methylamino ethyl)-carbamicacid tert-butyl ester (53 mg, 0.25 mmol), HBTU (140 mg, 0.38 mmol), andDIPEA (0.088 mL, 0.51 mmol). After 48 h stirring at room temperature,the resulting mixture is concentrated and diluted with saturated NaHCO₃solution and extracted with ethyl acetate (3×100 mL). The combineorganics are dried over Na₂CO₃ and concentrated to give a reddishresidue. Followed by deprotection using General Procedure A to give2-{5-[(2-amino-ethyl)-methyl-carbamoyl]-pyridin-2-ylamino}-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 77%). MS(ESI) m/z 451.3 (M+H)⁺

Example 67-Cyclopentyl-2-(4-dimethylamino-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (100 mg, 0.34 mmol) andN-4,N-4-dimethyl-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-4,6′-diamine(113 mg, 0.51 mmol) to give7-cyclopentyl-2-(4-dimethylamino-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (80 mg, 50%). MS(ESI) m/z 477.3 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.20 (1H, s), 8.74 (1H, s), 8.13 (1H, d),7.98 (1H, s), 7.43 (1H, d), 6.59 (1H, s), 4.80-4.68 (1H, m), 3.66 (2H,d), 3.10 (6H, s), 2.70-2.60 (2 h, m), 2.40-2.30 (2H, m), 2.20 (6H, s),2.00-2.80 (4H, m), 1.85-1.75 (2H, m), 1.70-1.60 (2H, m), 1.65-1.45 (2H,m).

Example 207-Cyclopentyl-2-(4-hydroxy-3,4,5,6-tetrahydro-2H-[1,3]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (290 mg, 0.939 mmol) and4-(tert-butyl-dimethyl-silanyloxy)-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamine(336 mg, 1.09 mmol). Followed by deprotection using 7 mL of TBAF in 28mL of THF to give7-cyclopentyl-2-(4-dimethylamino-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (110 mg, 61%). MS(ESI) m/z 450.3 (M+H)⁺

Example 357-Cyclopentyl-2-[4-(2-hydroxy-ethyl)-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (100 mg, 0.34 mmol) and2-(6′-amino-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-4-yl)-ethanol (90mg, 0.38 mmol) to give7-cyclopentyl-2-[4-(2-hydroxy-ethyl)-3,4,5,6-tetrahydro-2H-[1,3′]bipyridinyl-6′-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (80 mg, 93%). MS(ESI) m/z 478.3 (M+H)⁺

Example 527-Cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (300 mg, 1.03 mmol) and6-amino-3′,4′,5′,6′-tetrahydro-2′H-[3,4′]bipyridinyl-1′-carboxylic acidtert-butyl ester (313 mg, 1.13 mmol), followed by deprotection usingGeneral Procedure A to give7-cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (212 mg, 48%). MS(ESI) m/z 434.3 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.54 (1H, s), 8.80 (1H, s), 8.29 (1H, d),8.17 (1H, s), 7.62 (1H, d), 6.63 (1H, s), 4.83 (1H, m), 3.38-3.30 (3H,m), 3.06 (6H, s), 3.05-2.95 (1H, m), 2.88-2.80 (1H, m), 2.48-2.40 (4H,m), 2.04-1.95 (4H, m), 1.83-1.70 (2H, m), 1.70-1.64 (2H, m).

Example 807-Cyclopentyl-2-(1′-isopropyl-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a suspension of7-cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.23 mmol) in dichloromethane/acetone isadded NaBH(OAc)₃ (488 mg, 2.3 mmol) followed by 3 drops of glacialacetic acid. After reaction is completed and concentrated. Diluted with100 mL of H₂O and basified to pH12 with 50% NaOH solution dropwise (2mL). Extracted with dichloromethane (3×100 mL) and concentrated to give7-cyclopentyl-2-(1′-isopropyl-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (65 mg, 60%). MS(ESI) m/z 476.3 (M+H)⁺

Example 1007-Cyclopentyl-2-[1′-(2-hydroxy-ethyl)-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (63 mg, 0.15 mmol) and 2-bromo ethanol (90 mg, 0.72mmol) to give7-cyclopentyl-2-[1′-(2-hydroxy-ethyl)-1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (37 mg, 53%). MS(ESI) m/z 478.3 (M+H)⁺

Example 454-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridazin-3-yl]-piperazine-1-carboxylicacid tert-butyl ester

Following General Procedure D,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (200 mg, 0.68 mmol) and4-(6-amino-pyridazin-3-yl)-piperazine-1-carboxylic acid tert-butyl ester(210 mg, 0.75 mmol) to give4-[6-(7-cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridazin-3-yl]-piperazine-1-carboxylicacid tert-butyl ester (150 mg, 46%). MS(ESI) m/z 536.3 (M+H)⁺

Example 677-Cyclopentyl-2-(6-piperazin-1-yl-pyridazin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure A,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (150 mg, 0.28 mmol) gave7-cyclopentyl-2-(6-piperazin-1-yl-pyridazin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (2 mg, 2%). MS(ESI) m/z 436.3 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.77 (1H, s), 8.76 (1H, s), 8.20 (1H, d),7.39 (1H, d), 6.60 (1H, s), 5.75 (1H, s), 4.76-4.67 (1H, m), 3.52 (4H,s), 3.05 (6H, s), 2.94 (4H, s), 2.42-2.26 (2H, m), 1.97-1.88 (4H, m),1.62-1.56 (2H, m).

Example 707-Cyclopentyl-2-[6-(4-isopropyl-piperazin-1-yl)-pyridazin-3-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a suspension of7-cyclopentyl-2-(1′,2′,3′,4′,5′,6′-hexahydro-[3,4′]bipyridinyl-6-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.23 mmol) in dichloromethane/acetone isadded NaBH(OAc)₃ (488 mg, 2.3 mmol) followed by 3 drops of glacialacetic acid. After reaction is completed and concentrated. Diluted with100 mL of H₂O and basified to pH12 with 50% NaOH solution dropwise (2mL). Extracted with dichloromethane (3×100 mL), and concentrated to give7-cyclopentyl-2-[6-(4-isopropyl-piperazin-1-yl)-pyridazin-3-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (77 mg, 70%). MS(ESI) m/z 478.3 (M+H)⁺

Example 377-Cyclopentyl-2-{6-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridazin-3-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(6-piperazin-1-yl-pyridazin-3-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (100 mg, 0.229 mmol) and 2-bromo ethanol (143 mg,1.14 mmol) gave7-cyclopentyl-2-{6-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-pyridazin-3-ylamino}-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (14 mg, 13%). MS(ESI) m/z 480.3 (M+H)⁺

Example 487-Cyclopentyl-2-(3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (100 mg, 0.342 mmol) and5′-amino-2,3,5,6-tetrahydro-[1,2′]bipyrazinyl-4-carboxylic acidtert-butyl ester (114 mg, 0.408 mmol), followed by deprotection usingGeneral Procedure A to give7-cyclopentyl-2-(3,4,5,6-tetrahydro-2H-[1,2]bipyrazinyl-5′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (82 mg, 45%). MS(ESI) m/z 436.3 (M+H)⁺

Example 157-Cyclopentyl-2-[4-(2-hydroxy-ethyl)-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

Following General Procedure D,7-cyclopentyl-2-(3,4,5,6-tetrahydro-2H-[1,2]bipyrazinyl-5′-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (50 mg, 0.114 mmol) and 2-bromo ethanol (25 mg, 0.20mmol) to give7-cyclopentyl-2-[4-(2-hydroxy-ethyl)-3,4,5,6-tetrahydro-2H-[1,2′]bipyrazinyl-5′-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (30 mg, 54%). MS(ESI) m/z 480.6 (M+H)⁺

¹H NMR (400 MHz, CDCl₃): 9.19 (1H, s), 8.61 (1H, s), 7.76 (1H, s), 7.48(1H, s), 7.19 (1H, s), 6.36 (1H, s), 4.80-4.68 (1H, m), 3.66-3.57 (2 h,s), 3.54 (6H, s), 2.65 (3H, s), 2.59 (2 h, s), 2.56-2.40 (2H, m),2.03-1.93 (3 h, m), 1.68-1.56 (4H, m).

Example 407-(4-Hydroxy-4-methyl-cyclohexyl)-2-(pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

To a solution of7-(4-oxo-cyclohexyl)-2-(pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (25 mg, 0.066 mmol) in THF is added 20 drops ofMeMgI. After reaction is completed, added 25 mL of water then 30 mL ofaqueous sodium bicarbonate. Extraction with dichloromethane (3×50 mL)and concentrated to give a mixture of diastereomers. Preparative HPLC togive7-(4-hydroxy-4-methyl-cyclohexyl)-2-(pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (2 mg, 4%). MS(ESI) m/z 395.3 (M+H)⁺

Example 587-Cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid methylamide

Following Buchwald Method B,2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acidmethylamide (500 mg, 1.80 mmol) and4-(6-amino-pyridin-3-yl)-piperazine-1-carboxylic acid tert-butyl ester(550 mg, 1.98 mmol) gave7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid methylamide (580 mg, 77%). MS(ESI) m/z 421.2 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 8.72 (1H, s), 8.38 (1H, d), 8.02 (1H, s),7.77 (1H, s), 7.36 (1H, dd), 6.67 (1H, s), 6.16-6.10 (1H, m), 5.50-5.48(1H, m), 3.15 (3H, d), 3.03 (2H, d), 2.68-2.58 (2H, m), 2.14-2.05 (4H,m), 1.80-1.61 (8H, m).

Example 517-Cyclopentyl-2-[5-(4-isopropyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid methylamide

To a suspension of2-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acidmethylamide (500 mg, 1.20 mmol) in acetone is added NaBH(OAc)₃ (2.5 g,12 mmol) followed by 15 drops of glacial acetic acid. After reaction iscompleted and concentrated. Diluted with 250 mL of H₂O and basified topH12 with 50% NaOH solution dropwise. Extracted with dichloromethane(3×250 mL) and concentrated to give7-cyclopentyl-2-[5-(4-isopropyl-piperazin-1-yl)-pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid methylamide (277 mg, 50%). MS(ESI) m/z 463.4 (M+H)⁺

Example 11(7-Cyclopentyl-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-yl-pyridin-2-yl)-amine

Following Buchwald Method B,2-chloro-7-cyclopentyl-6-oxazol-5-y-l-7H-pyrrolo[2,3-d]pyrimidine (70mg, 0.24 mmol) and 4-(6-amino-pyridin-3-yl)-piperazine-1-carboxylic acidtert-butyl ester (74 mg, 0.27 mmol), followed by deprotection usingGeneral Procedure A to give(7-cyclopentyl-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-yl-pyridin-2-yl)-amine(25 mg, 24%). MS(ESI) m/z 431.2 (M+H)⁺

¹H NMR (400 MHz, DMSO-d6): 9.33 (1H, s), 8.78 (1H, s), 8.59 (1H, s),8.13 (1H, d), 7.98 (1H, d), 7.61 (1H, s), 7.40 (1H, dd), 6.78 (1H, s),4.70-4.77 (1H, m), 3.04-3.01 (4H, m), 2.86-2.84 (4H, m), 2.03-2.01 (6H,m), 1.68-1.67 (2H).

Example 18(7-Cyclopentyl-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-ylmethyl-pyridin-2-yl)-amine

Following Buchwald Method B,2-chloro-7-cyclopentyl-6-oxazol-5-y-l-7H-pyrrolo[2,3-d]pyrimidine (100mg, 0.346 mmol) and4-(6-amino-pyridin-3-ylmethyl)-piperazine-1-carboxylic acid tert-butylester (106 mg, 0.363 mmol), followed by deprotection using GeneralProcedure A to give(7-cyclopentyl-6-oxazol-5-yl-7H-pyrrolo[2,3-d]pyrimidin-2-yl)-(5-piperazin-1-ylmethyl-pyridin-2-yl)-amine(23 mg, 15%). MS(ESI) m/z 445.2 (M+H)⁺

Example 1097-cyclopentyl-2-[5-(2-oxopiperazin-1-yl)pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

A mixture of 5-bromo-2-nitropyridine (200 mg, 1 mmol),1-Boc-3-oxopiperazine (240 mg, 1.2 mmol), Xantphos (43 mg, 0.075 mmol),cesium carbonate (326 mg, 1 mmol), palladium(II) acetate (11 mg, 0.049mmol) in dioxane (5.5 mL) is heated to 120° C. in a Personal Chemistrymicrowave apparatus for 0.5 h. TLC and LCMS analysis indicatescompletion of the reaction. The reaction mixture is filtered throughCelite, evaporated in vacuo, and the residue is partitioned betweenwater and ethyl acetate. The organic layer is washed with brine, dried(Na2SO4) and evaporated in vacuo. Purification by flash chromatographyon silica (ethyl acetate) provides4-(6-Nitropyridin-3-yl)-3-oxopiperazine-1-carboxylic acid tert-butylester as a pale brown solid (248 mg, 77%). MS (ESI) m/z 323 [M+H+].

By repeating procedures described in Example B,4-(6-nitropyridin-3-yl)-3-oxopiperazine-1-carboxylic acid tert-butylester (240 mg, 0.74 mmol) gives4-(6-aminopyridin-3-yl)-3-oxopiperazine-1-carboxylic acid tert-butylester (225 mg). MS (ESI) m/z 293 [M+H]+.

A mixture of2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (225 mg, 0.77 mmol),4-(6-aminopyridin-3-yl)-3-oxopiperazine-1-carboxylic acid tert-butylester (204 mg, 0.69 mmol), BINAP (24 mg, 0.038 mmol), palladium(II)acetate (6 mg, 0.027 mmol) and cesium carbonate (340 mg, 1.05 mmol) indioxin (4 mL) is flushed with nitrogen and heated to 100° C. overnight.Additional palladium(II) acetate (6 mg, 0.027 mmol) and BINAP (24 mg,0.038) are added and heating is continued at 110° C. for 2 h at whichpoint LCMS and TLC analysis indicates completion of the reaction. Thesolvent is removed in vacuo and the residue is stirred in water withsonication in an ultrasonic bath. The suspension is filtered and thefilter cake is washed with heptane. Drying in vacuo provides4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-3-oxo-piperazine-1-carboxylicacid tert-butyl ester as a tan solid (350 mg, 83%) MS (ESI) m/z=549[M+H]+.

Following General Procedure A4-[6-(7-Cyclopentyl-6-dimethylcarbamoyl-7H-pyrrolo[2,3-d]pyrimidin-2-ylamino)-pyridin-3-yl]-3-oxo-piperazine-1-carboxylicacid tert-butyl ester gives7-cyclopentyl-2-[5-(2-oxopiperazin-1-yl)pyridin-2-ylamino]-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide (10 mg, 3.5%) MS (ESI) m/z=448 [M+H]+.

Example 1107-Cyclopentyl-2-(5-morpholin-4-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide

A mixture of 5-Morpholin-4-yl-pyridin-2-ylamine (0.61 g, 3.4 mmol;prepared using methods similar to those described in Example A andExample B),2-Chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic aciddimethylamide (1.00 g, 3.4 mmol), BINAP (106 mg, 0.17 mmol),palladium(II) acetate (38 mg, 0.17 mmol) and cesium carbonate (1.6 g,4.9 mmol) in dioxan (20 mL) is heated to 110° C. for 6 h. After coolingto room temperature, heptane (30 mL) is added and the mixture is stirredfor 1 h. The resulting suspension is filtered and the filtercake issuspended in water with vigorous stirring. The resulting suspension isagain filtered and the filtercake is washed with water then diethyletherbefore being dried in vacuo to provide7-Cyclopentyl-2-(5-morpholin-4-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylicacid dimethylamide as a tan solid (1.30 g, 88%) MS (ESI) m/z=436.1[M+H]+.

The following tables 1 and 2 of compounds are examples of compoundswhich may be made using the synthetic routes exemplified in theexperimental section. While the synthesis of all compounds is not shown,one of skill in the art may be able to make each compound using thesynthetic routes shown.

TABLE 1 Example Compound Number

 1

 2

 3

 4

 5

 6

 7

 8

 9

10

   10A

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100 

101 

102 

103 

104 

105 

106 

107 

108 

109 

110 

Biological Activity CDK4/cyclin D1 Enzymatic Activity Assay

A 384-well microtiter Lance TR-FRET (time-resolved-fluorescence energytransfer) endpoint assay was used for CDK4/cyclin D1 kinase activitymeasurements. The same assay was used for IC50 determination of smallmolecule inhibitors. In general, the kinase reactions were carried outin 30 μL volumes in the reaction solution containing the following: 2 uLcompound (in 20% DMSO), 18 uL CDK4/cyclin D1 in Assay Buffer (50 mMHEPES, pH 7.5, 5 mM MgCl₂, 2 mM MnCl₂, 1 mM DTT, 0.05% BSA, 0.02%Tween-20), 10 uL of the mixture of pRb152 and ATP. The final reactionmixture contains compound (inhibitor) with the concentration varyingfrom 0.005-10 μM, 2% DMSO, 0.3 nM CDK4/cyclin D1, 175 nM pRb152, and 3μM ATP (Amersham Pharmacia, Cat. No. 27-2056-01). All reactions were runat room temperature in 384-well white flat-bottom OptiPlates (PerkinElmer, Cat. No. 6007290) for 60 min then were quenched by the additionof 10 μL of 120 mM EDTA. The signals were captured by the addition of 40μL of the Detection Solution containing the following: Detection Buffer(50 mM HEPES, pH 7.5, 30 mM EDTA, 0.1% Triton x-100, 0.05% BSA), 70ng/mL anti-phospho-pRb(5780) (Cell Signaling Technology, Cat. No.9307S), 1 nM Lance Eu-W1024-Rabbit anti-IgG (Perkin Elmer, Cat. No.AD0082), and 20 nM SureLight™ Allophycocyanin-Streptavidin (PerkinElmer, Cat. No. CR130-100). The resulted solutions were incubated atroom temperature for 2 hours before read on the Evision MultilabelReader (Perkin Elmer, Envision 2102-0010). Note: IC₅₀<0.005 nM orIC₅₀>10 μM indicates the true IC₅₀ is out of detection range.

CDk4/cyclin D1 recombinant protein used in the enzymatic activity assaywas prepared by coexpressing pDEST10-CDK4 (N-terminal His₆) andpFastBacDual-GST-hCyclinD1 viruses in Sf21 cells. The overexpressedprotein was purified by Ni-NTA affinity pull down to >80% pure by SizingHPLC.

CDK1/cyclin B Enzymatic Activity Assay

A 384-well microtiter IMAP-FP™ (Molecular Devices Trade Mark Technology)endpoint assay was used for CDK1/cyclin B kinase activity measurements.The same assay was used for IC₅₀ determination of small moleculeinhibitors. In general, the kinase reactions were carried out in 20 μLvolumes in the reaction solution, which is composed of 2 μL compound (in20% DMSO), 8 μL CDK1/cyclin B in the 1× Reaction Buffer (MolecularDevices, Cat. No. R8139), 10 μL substrate mixture of Tamra Histone-H1peptide (Molecular Devices, Cat. No. R7384) and ATP (Amersham Pharmacia,Cat. No. 27-2056-01) in the 1× Reaction Buffer with 1 mM DTT freshlyadded. The final reaction mixture contains compound (inhibitor) with theconcentration varying from 0.005-10 μM, 2% DMSO, 0.25 nM CDK1/cyclin B,100 nM Tamra Histone-H1 peptide, and 20 μM ATP.

All reactions were run at room temperature in black 384-well flat-bottomCostar plates (Corning, Cat. No. 3710) for 120 min then were quenched bythe addition of 60 μL 400-fold diluted 1× Progressive Binding Buffer A(Molecular Devices, Cat. No. R8139). The fluorescent polarizationsignals were read on the Evision Multilabel Reader (Perkin Elmer,Envision 2102-0010) after 2-hour incubation at room temperature. Note:IC₅₀<0.005 nM or IC₅₀>10 μM indicates the true IC₅₀ is out of detectionrange.

CDK2/Cyclin a Enzymatic Activity Assay

The assay was run under the conditions identical to that for CDK1/cyclinB except 0.25 nM CDK1/cyclin B was replaced with 0.3 nM CDK2/cyclin A.The results of the assays are summarized in table 2.

TABLE 2 Example No. CDK4 (μM) CDK1 (μM) CDK2 (μM) MS (MH+) 1 * >15 >15351.1 74 *** >15 >15 435.3 78 *** >15 >15 463.3 86 ** >15 >15 481.3 26** >15 >15 479.3 14 ** >15 >15 493.3 95 ** >15 >15 521.3 33 ** >15 >15523.4 57 ** >15 >15 493.3 56 ** >15 >15 493.3 71 ** >15 >15 507.3 21*** >15 >15 493.3 44 ** >15 >15 509.3 46 ** >15 >15 509.3 29 ** 15 >15509.3 79 ** >15 >15 503.3 63 *** >15 >15 477.3 36 ** >15 >15 493.4101 * >15 >15 521.4 103 ** >15 >15 507.3 69 ** >15 >15 531.4 92*** >15 >15 491.3 99 ** >15 >15 505.3 90 ** >15 >15 519.4 68 *** >15 >15519.4 25 ** >15 >15 517.3 10 ** >15 >15 492.4 84 ** >15 >15 474.3 9** >15 >15 488.3 7 ** >15 >15 477.3 27 ** >15 >15 503.3 23 ** >15 >15545.3 90 ** >15 >15 559.4 91 ** >15 >15 559.4 12 ** >15 >15 492.3 88*** >15 >15 520.5 22 ** >15 >15 532.3 94 ** >15 >15 546.3 38 ** >15 >15548.3 30 * >15 >15 549.3 31 * >15 >15 549.3 19 *** >15 >15 448.3 16*** >15 >15 449.3 81 ** >15 >15 493.3 17 ** >15 >15 449.3 82 ** >15 >15493.3 72 ** >15 >15 463.3 24 ** >15 >15 463.3 4 ** >15 >15 449.2 8 ** 148 449.3 13 ** >15 >15 435.3 39 ** >15 >15 436.3 32 ** >15 >15 422.5 59** >15 >15 423.4 83 * >15 >15 424.2 10A ** 12 14 436.3 34 * >15 >15527.4 42 ** >15 >15 449.3 43 ** >15 >15 463.6 54 ** >15 >15 463.3 55** >15 >15 463.4 60 * >15 >15 477.4 61 * >15 >15 491.5 62 ** >15 >15491.4 65 ** >15 >15 506.4 73 ** >15 >15 475.6 75 ** >15 >15 477.2 76** >15 >15 464.4 77 ** >15 >15 507.5 85 ** >15 >15 493.4 66 ** >15 >15438.3 3 ** >15 >15 477.3 53 ** >15 >15 491.3 49 ** >15 >15 463.3 96** >15 >15 507.3 50 ** >15 >15 505.5 87 ** 13 >15 478.3 41 *** 14.8 4.7451.3 6 *** >15 >15 477.3 20 ** >15 >15 450.3 35 ** 4.7 2.9 478.3 52** >15 >15 434.3 80 ** >15 >15 476.3 100 ** >15 >15 478.3 45 * >15 20536.3 67 ** >15 >15 436.3 70 ** >15 >15 478.3 37 ** >15 >15 480.348 * >15 >15 436.3 15 * >15 >15 480.6 40 * >15 >15 395.3 47 *** 2.6 8.3431.3 58 ** >15 >15 421.2 51 ** >15 >15 463.4 11 *** 1.3 3.5 431.2 18*** 1.1 2.8 445.2 109 * >15 >15 448.5 110 * >15 >15 436.1 Key Greaterthan 0.1, and less than or equal to 1.0 = * Greater than 0.01, and lessthan or equal to 0.1 = ** Greater than 0.001, and less than or equal to0.01 = ***

1. A compound of formula I:

or a pharmaceutically acceptable salt, wherein X is CR⁹, or N; R¹ isC₁₋₈ alkyl, CN, C(O)OR⁴ or CONR⁵R⁶, a 5-14 membered heteroaryl group, ora 3-14 membered cycloheteroalkyl group; R² is C₁₋₈alkyl, C₃₋₁₄cycloalkyl, or a 5-14 membered heteroaryl group, and wherein R² may besubstituted with one or more C₁₋₈ alkyl, or OH; L is a bond,C₁₋₈alkylene, C(O), or C(O)NR¹⁰, and wherein L may be substituted orunsubstituted; Y is H, R¹¹, NR¹²R¹³, OH, or Y is part of the followinggroup

where Y is CR⁹ or N; where 0-3 R⁸ may be present, and R⁸ is C₁₋₈alkyl,oxo, halogen, or two or more R⁸ may form a bridged alkyl group; W isCR⁹, or N; R³ is H, C₁₋₈alkylR¹⁴, C₃₋₁₄cycloalkyl, C(O)C₁₋₈ alkyl, C₁₋₈haloalkyl, C₁₋₈alkylOH, C(O)NR¹⁴R¹⁵, C₁₋₈cyanoalkyl, C(O)R¹⁴,C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵, C₀₋₈alkylC(O)OR¹⁴, NR¹⁴R¹⁵, SO₂C₁₋₈alkyl,C₁₋₈alkylC₃₋₁₄cycloalkyl, C(O)C₁₋₈alkylC₃₋₁₄cycloalkyl, C₁₋₈alkoxy, orOH which may be substituted or unsubstituted when R³ is not H. R⁹ is Hor halogen; R⁴, R⁵, R⁶, R⁷, R¹⁰, R¹¹, R¹², R¹³, R¹⁴, and R¹⁵ are eachindependently selected from H, C₁₋₈alkyl, C₃₋₁₄ cycloalkyl, a 3-14membered cycloheteroalkyl group, a C₆₋₁₄ aryl group, a 5-14 memberedheteroaryl group, alkoxy, C(O)H, C(N)OH, C(N)OCH₃, C(O)C₁₋₃alkyl,C₁₋₈alkylNH₂, C₁₋₆ alkylOH, and wherein R⁴, R⁵, R⁶, R⁷, R¹⁰, R¹¹, R¹²,and R¹³, R¹⁴, and R¹⁵ when not H may be substituted or unsubstituted; mand n are independently 0-2; and Wherein L, R³, R⁴, R⁵, R⁶, R⁷, R¹⁰,R¹¹, R¹², and R¹³, R¹⁴, and R¹⁵ may be substituted with one or more ofC₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl, C₃₋₁₄cycloalkyl, 5-14 memberedheteroaryl group, C₆₋₁₄aryl group, a 3-14 membered cycloheteroalkylgroup, OH, (O), CN, alkoxy, halogen, or NH₂.
 2. The compound of formulaI according to claim 1 wherein R³ is H, C₁₋₈alkyl, C₃₋₁₄cycloalkyl,C(O)C₁₋₈ alkyl, C₁₋₈alkylOH, C₁₋₈cyanoalkyl,C₀₋₈alkylC(O)C₀₋₈alkylNR¹⁴R¹⁵, C₀₋₈alkylC(O)OR¹⁴, NR¹⁴R¹⁵,C₁₋₈alkylC₃₋₁₄cycloalkyl, C(O)C₁₋₈alkylC₃₋₁₄cycloalkyl, C₀₋₈alkoxy,C₁₋₈alkylR¹⁴, C₁₋₈haloalkyl, or C(O)R¹⁴, which may be substituted withone or more of OH, CN, F, or NH₂, and wherein R¹⁴ and R¹⁵ are eachindependently selected from H, C₁₋₈alkyl, C₃₋₁₄cycloalkyl, alkoxy,C(O)C₁₋₃alkyl, C₁₋₈alkylNH₂, or C₁₋₆alkylOH.
 3. The compound of formulaI according to claim 1 wherein R³ is H, C₁₋₈alkyl, or C₁₋₈alkylOH. 4.The compound of formula I according to claim 1 wherein Y is H, OH, or Yis part of the following group

where Y is N and W is CR⁹, or N; where 0-2 R⁸ may be present, and R⁸ isC₁₋₈alkyl, oxo, or two or more R⁸ may form a bridged alkyl group.
 5. Thecompound of formula I according to claim 1 wherein L is a bond,C₁₋₈alkylene, or C(O)NH, or C(O).
 6. The compound of formula I accordingto claim 1 wherein R² is C₃₋₁₄cycloalkyl.
 7. The compound of formula Iaccording to claim 1 wherein R² is cyclopentane.
 8. The compound offormula I according to claim 1 wherein R¹ is CN, C(O)OR⁴, CONR⁵R⁶, or a5-14 membered heteroaryl group.
 9. The compound of formula I accordingto claim 1 wherein R¹ is CONR⁵R⁶, and R⁵ and R⁶ are C₁₋₈alkyl.
 10. Thecompound of formula I according to claim 1 wherein X is CR⁹.
 11. Thecompound of formula I according to claim 1 wherein one X is N and theother X is CR⁹.
 12. The compound of formula I according to claim 1wherein X is CR⁹ and Y is

where m and n are 1, and Y and W are N.
 13. A method of treating adisease, disorder or syndrome associated with CDK 4 inhibition, saidmethod comprising administering a compound according to claim 1 or itsprodrug or pharmaceutical composition comprising the compound of formulaI or its prodrug and pharmaceutically acceptable excipients to a subjectin need thereof.
 14. The method of treating as claimed in claim 13,wherein the disease, disorder or syndrome is hyperproliferative in asubject, wherein subject is an animal including humans, selected from agroup comprising cancer and inflammation.
 15. A method of inhibiting acyclin dependent kinase (e.g. cdk-4), which method comprises contactingthe kinase with a kinase-inhibiting compound according to claim
 1. 16. Amethod of modulating a cellular process (for example cell division) byinhibiting the activity of a cyclin dependent kinase using a compoundaccording to claim
 1. 17.-18. (canceled)
 19. A pharmaceuticalcomposition comprising a compound according to claim 1 and apharmaceutically acceptable carrier.
 20. A pharmaceutical compositioncomprising a compound according to claim 1 and a pharmaceuticallyacceptable carrier in a form suitable for oral administration.
 21. Acompound of formula I(a):

or a pharmaceutically acceptable salt thereof, wherein: R⁵⁰ isCONR⁵⁴R⁵⁵, or CN; R⁵¹ is C₃₋₁₄cycloalkyl which may be unsubstituted orsubstituted by C₁₋₃alkyl, or OH; Z is CH or N; and V is NR⁵⁶ or CHR⁵⁷;R⁵⁴ and R⁵⁵ are independently H, C₁₋₃ alkyl, R⁵², R⁵³ R⁵⁶, and R⁵⁷ areindependently H, C₁₋₈alkyl, C₃₋₁₄cycloalkyl, C₁₋₈haloalkyl, NR⁵⁸R⁵⁹,C(O)OR⁶⁰, C(O)C₁₋₈alkyl, C₀₋₈alkylC(O)C₀₋₈alkyl-NR⁶¹R⁶², C₁₋₈alkoxy,C₁₋₈alkylOR⁶³, C(O)-5-14cycloheteroalkyl group, C₃₋₁₄cycloalkyl group,each of which when not H may be substituted by one or more of C₁₋₈ alky,OH, or CN; R⁵⁸, R⁵⁹, R⁶⁰, R⁶¹, R⁶², and R⁶³ are H or C₁₋₈alkyl. 22.(canceled)